JP2003063974A - Antioxidant, nitrogen monooxide-production inhibitor, antiulcer drug and food containing the drug component - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an antiulcer drug having broad medicinal effectiveness which is derived from a natural plant and has anti-oxidation activity and nitrogen monoxide production-inhibiting activity. SOLUTION: An antioxidant, nitrogen monoxide-production inhibitor or antiulcer drug which contains as the active ingredients one or more plants selected from the group consisting of Salacia chinensis, Salacia oblonga and Salacia reticulate or their extracts, and processed food which contains the drug components.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an antioxidant, a nitric oxide production inhibitor, an antiulcer agent, which contains a natural plant-derived composition as an active ingredient, and a processed food containing the drug ingredient.

[0002]

2. Description of the Related Art In recent years, it has been found that active oxygen and lipid peroxide have a close relationship with gastrointestinal mucosal diseases, and oral ingestion of antioxidants is a treatment for gastrointestinal ulcer disease. Or, it has become clear that it is effective for prevention.

[0003] Conventionally, a large number of compositions derived from chemical compounds or natural products have been proposed as antioxidants for the purpose of suppressing the oxidation of foods and eliminating free radicals in the living body.
Since a synthetic composition is likely to have side effects, it is strongly desired to use a natural composition derived from a natural product, which has a mild antioxidant effect and has no side effects even after long-term ingestion.

Therefore, various compositions mainly derived from natural products have been proposed, which have both an antioxidant effect and an effect as an anti-ulcer agent. For example, "food and drink for preventing gastric ulcer containing antioxidants extracted from cacao beans"
(JP-A-7-274894), "Antioxidant composition comprising natto bacteria, lactic acid bacteria and the like and anti-ulcer agent" (JP-A-9-
154535), "Pharmaceuticals and antioxidants containing a novel isoflavone derivative or a salt thereof as an active ingredient, which is mainly collected from a culture of a microorganism" (JP-A-10-599).
No. 56) or “N mainly extracted from sugar beet seeds, N
-Trans-feruloyl homovanillylamine and the like as active ingredients, plant antioxidants and therapeutic agents for gastric ulcers and the like "(JP-A 2000-129257).

On the other hand, in the living body, nitric oxide (hereinafter sometimes abbreviated as NO) is circulated in the circulatory system, nervous system,
It has become known that it has various actions as a physiologically active substance such as the immune system, and it has become clear that NO is closely related to various inflammations.

In vivo, NO is produced by oxidizing L-arginine by nitric oxide synthase (NOS). There are non-inducible and inducible isozymes in NOS, inducible NOS (iNOS) is macrophage,
Although abundant in endothelial cells, smooth muscle cells, etc., it has been revealed that a therapeutic effect can be obtained in various inflammations by inhibiting the activity of iNOS and suppressing the production of NO.

For this reason, a number of synthetic and natural products-derived nitric oxide production inhibitors have been proposed in recent years. Examples of the natural products include, for example, "anti-inflammatory compounds containing a polyacetylene compound, which is a component containing Hyugatouki. Composition such as an agent "(Japanese Patent Laid-Open No. 11-310527) and" Scientific name And
A nitric oxide production inhibitor containing at least one plant selected from the group consisting of whole plants of rographis paniculata or an extract thereof as an active ingredient "(JP 2000-34A).
No. 233) and the like.

[0008]

Therefore, the present invention is derived from a natural plant which has no side effects even if taken for a long period of time, and has an antioxidant action and a nitric oxide production inhibitory action, and is an anti-ulcer agent. The purpose of the present invention is to provide a drug having a wide range of therapeutic effects.

[0009]

[Means for Solving the Problems] As a result of intensive studies to solve the above problems, the present inventors have found that plants belonging to the genus Salacia belonging to the family Euonymus or Dechinmulaceae, particularly Salacia chinensis, Salacia Salacia oblonga, Salacia reticulata
It has been found that (Salacia reticulata) contains a component exhibiting excellent medicinal effects in antioxidant action, NO production inhibitory action and antiulcer action. These plants have been used as crude drugs since ancient times in Indian or Thai traditional medicine, and it is guaranteed that there will be no side effects even if they are taken for a long period of time.

The present invention based on this finding is an antioxidant containing, as an active ingredient, one or more plants selected from the group consisting of Salacia quinensis, Salacia oblonga and Salacia reticulata.

In addition, Salacia kinensis, Salacia
It is a nitric oxide production inhibitor containing, as an active ingredient, one or more plants selected from the group consisting of oblonga and Salacia reticulata or an extract therefrom.

In addition, Salacia kinensis, Salacia
It is an anti-ulcer agent containing, as an active ingredient, one or more plants selected from the group consisting of oblonga and Salacia reticulata or an extract therefrom.

Further, it is a processed food containing one or more plant powders selected from the group consisting of Salacia quinensis, Salacia oblonga and Salacia reticulata or an extract thereof.

The above "extract" may be one extracted from one of the above plant groups, a mixture of two or more kinds of these extracts, or the above. It may be one extracted from two or more kinds of plant groups at the same time.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION Salacia quinensis (hereinafter referred to as S
Abbreviated as C) is a vine shrub (height 5 m or less) that grows naturally in Southeast Asia (Thailand, Malaysia, Southern China, etc.), India, Sri Lanka, etc., and its leaves are oval with a length of 4-8 cm, yellow. Or bloom green-yellow flowers and turn red when ripe 1
It is a plant with ~ 2 cm of small spherical fruit.

Salacia oblonga (hereinafter abbreviated as SO) and Salacia reticulata (hereinafter abbreviated as SR) are often found in India, Sri Lanka and the like,
It is a plant similar to SC, but the tree height is lower than SC,
Actually, the difference is 3 to 5 cm, which is larger than SC. S
All of C, SO, and SR have been widely used since ancient times as crude drugs having various medicinal effects in traditional medicine such as Thailand and India.

In the present invention, any of the above plants (bark, wood, leaves, etc.) and underground (root bark, rhizome) can be used, but it contains many medicinal components. From this point of view, it is preferable to use the bark or the wood part (hereinafter, the description about the embodiment of the invention is common to any of SC, SO, and SR unless otherwise specified). Alternatively, the plant itself or a dried and crushed plant may be used, or an extract extracted from this with water or an organic solvent may be used.

Examples of the organic solvent include, but are not limited to, lower alcohols such as ethanol, methanol, propanol and butanol, and esters such as ethyl acetate. From the viewpoint of economy and safety, extraction with water and / or ethanol is preferable.

In the case of extraction with water, a dried product of a plant or a crushed product thereof is heated to about 2 to 5 times its volume (60 to 9).
It may be extracted by immersing in (0 ° C.) for about 1 to 5 hours. It may be extracted by immersing it in water at room temperature for a long time (1 to 5 days). The extract or its concentrated extract may be taken as it is, but usually a dry powder thereof is used.

In the case of extraction with ethanol, the plant or its pulverized product is heated at room temperature or low temperature (2 to 5 times) (3
It may be extracted by immersing it in ethanol (0 to 80 ° C.) for about 1 to 5 hours. The extract is at a relatively low temperature (50 ° C. or lower), and if necessary, ethanol can be volatilized under reduced pressure to obtain a dry powder. The yield of the extract (dry weight of extract / dry weight of plant x 100) is 15 to 15 in the case of water extraction.
It is about 25%, and in the case of ethanol extraction, it is about 25 to 35%.

In any of the cases where the agent of the present invention is used as an antioxidant, a nitric oxide production inhibitor or an antiulcer agent, a dry powder of a crushed plant product or an extract may be taken as it is. , Add excipients (lactose, starch, dextrin, gelatin, celluloses, etc.) to it,
It may be taken as a solid preparation such as tablets, granules and capsules. Further, a pulverized product of the plant, a dry powder of the extract or a concentrated extract may be taken as a tea preparation.

The oral dose may be about 1 to 500 mg / kg body weight of the extract per dry dose, and in the case of a pulverized plant, it may be about 5 times this amount. The number of doses is not particularly limited, but may be 1 to 5 times a day. From the viewpoint of prevention or treatment of ulcers of the digestive system, it is preferable to take it about 3 times a day after eating.

Further, in the present invention, the pulverized product or extract of the above plant may be contained in various processed foods and orally ingested. Examples of processed foods used for this purpose include processed foods containing carbohydrate as a main component (breads, noodles, etc.), processed fish meat products, processed dairy products, and various confectionery products.

In the above-mentioned food processing process, particularly in the raw material blending process, by adding an appropriate amount of the above-mentioned plant ground product or extract and mixing and stirring to adjust the raw materials,
A processed food containing the drug of the present invention can be obtained in the same manner as in the usual manufacturing process. The amount to be added may be appropriately selected so as to substitute a part thereof within the range that does not deteriorate the flavor of food, using the above oral dose as a guide.

There are various actions of NO in the living body, for example, vasorelaxant action, platelet aggregation inhibitory action, leukocyte adhesion / migration inhibitory action, sympathetic nerve activity inhibitory action, action as a signal transmitter between nerve cells and the like. Although it has various physiological actions, it acts as an obstacle when NO is excessively produced in vivo. Therefore, the nitric oxide production inhibitor of the present invention is not only an anti-inflammatory agent, but also, for example, septic shock, hypotension, ischemic disease, allergic disease, autoimmune disease, rheumatoid arthritis, insulin-dependent diabetes mellitus. A wide range of medicinal effects are expected for the prevention or treatment of the above.

[0026]

[Examples] As test substances of Examples, SC, SO and SR extracts prepared by the following method were used, and DPPH (1,1-diphenyl-2-picryl hydraz) as an antioxidant activity was used.
yl) Radical and superoxide scavenging activity, macrophage-derived nitric oxide production inhibitory activity, and effect as an anti-ulcer agent were investigated by the following evaluation methods.

Method for preparing test substance Dry aerial parts of SC, SO and SR were crushed to 100 mesh or less, and 100 g of the crushed product was about 1 for each.
The extract was obtained by immersing it in liter of 70% ethanol. After keeping the temperature of ethanol at 70 to 80 ° C. and extracting for 3 hours, the extract was filtered, and the filtrate was evaporated at 30 ° C. under reduced pressure to evaporate ethanol to obtain an extract powder. Extraction rate is 15 to 25% (extract powder weight / crushed material weight x 1
00). The extract powder was further dissolved in dimethylsulfoxide (DMSO) to prepare an extract having a predetermined concentration, which was used as a test substance.

Evaluation method DPPH radical scavenging action: 4
60% ethanol / 4 containing 0 μM DPPH radical
A 0 mM acetate buffer (pH 5.5) was prepared, and this was left at room temperature for 30 minutes to allow the test substance to undergo a DPPH radical scavenging reaction, and then the absorbance at 517 nm was measured (Beckman DU530). Then, the amount of DPPH radicals remaining was determined. The above-mentioned measurement is performed by changing the concentration of the test substance in several steps, and the elimination amount becomes 50%. 5
The 0% capture concentration SC ₅₀ (μg of extract powder / ml of buffer) was determined.

Superoxide scavenging action: test substance at a predetermined concentration, 100 μM xanthine, 100 μM ED
TA, 0.005% bovine serum albumin, 25 μM N
BT (nitroblue tetrazolium) and 0.015
A 40 mM sodium carbonate buffer solution (pH 10.2) containing 3 U / ml xanthine oxidase was prepared and kept at 25 ° C. for 20 minutes to promote the reaction of superoxide generation and its elimination by the test substance. Then, 6 mM of copper chloride reagent was added for color development, and the absorbance at 560 nm was measured (Beckman DU530) to determine the amount of residual superoxide. The above-mentioned measurement was carried out by changing the concentration of the test substance in several stages, and the 50% trapping concentration SC ₅₀ (extract powder μg /
Buffer ml) was determined.

Macrophage-derived nitric oxide production inhibitory activity ddY male mice (body weight: around 30 g) were cervical dislocated,
Intraperitoneally 5-6 ml of phosphate buffered saline (137 m
MNaCl, 2.7 mM KCl, 1.5 mM KH ₂ P
O ₄ , 8.1 mM Na ₂ HPO ₄ , pH 7.4),
The washing solution was collected to obtain macrophages. This is RPM
I1640 medium (containing 5% fetal bovine serum, 100 U / ml penicillin, 100 μg / ml streptomycin)
Suspend the cells in a 96-well microplate (5 x 10
⁵ cells / well / 200 μl).

After culturing for 1 hour, the test substance at a predetermined concentration
And lipopox that stimulates macrophages to produce NO
R containing 10 μg / ml of saccharide (LPS)
It was transferred to PMI1640 medium and cultured for 20 hours. So
NO accumulated in the medium after ^2-The amount of grease reagent (58
mM Sulfanilamide, 3.9 mM N-1 naphthyle
Absorbed by coloring with dilendiamine (255 mM phosphoric acid)
Degree (Model 550 microplate reader, Biorad,
Measurement wavelength: 570 nm, reference wavelength: 655 nm), monoacid
The amount of nitrogen oxide (NO) produced was determined. Also, as a reference value,
With or without LPS containing no test substance
If NO, calculate NO production by the same method as above.
It was

Inhibitory effect on ethanol-induced gastric mucosal damage SD rats (body weight: around 250 g) were fasted for 24 hours and then suspended in a 5% gum arabic solution to be tested substances 0 to 2
00 mg / kg was orally administered, and 1 hour later, 1.5 ml of 99.5% ethanol was orally administered per rat.
One hour after administration, the stomach was extracted, fixed with 1.5% formalin, the length of the damaged site (Lesion index, mm) was determined, and the inhibition rate was calculated.

In the above evaluation methods, rats and mice were obtained from the Kiwa Laboratory for Animal Research and LP
S and RPMI1640 medium were purchased from Sigma, and other reagents were purchased from Wako Pure Chemical Industries, Ltd. using special grade reagents. Moreover, although the above-mentioned evaluation was performed using SC, SO, and SR as test substances for-, only SC was evaluated.

In Table 1, for SC, SO and SR,
The test substance concentration SC ₅₀ for eliminating 50% of DPPH radical or superoxide is shown. From these results, it is considered that the antioxidant of the present invention has an antioxidant activity equivalent to or higher than that of green tea or polyphenol.

[0035]

[Table 1]

In Table 2, for SC, SO and SR,
The evaluation result of the macrophage origin nitric oxide production inhibitory activity is shown. In the table, the test substance concentration is 3, respectively.
Mean value and variance of NO production inhibitory rate in the case of 10, 30, 100, 300 (extract powder µg / buffer solution ml), 50% NO production inhibitory concentration (µg obtained from these data
/ Ml) and the reference value NO when no test substance is added
The production amount (with and without addition of LPS) is shown.

From these results, it is considered that the nitric oxide production inhibitor of the present invention has a NO production inhibitory effect equivalent to or higher than that of conventional natural plant-derived agents. In addition, the symbol ▼ in the table indicates the case where cell damage is observed, and
For O and SR, if the concentration of the test substance becomes excessive, there is concern about cytotoxicity, so it is necessary to consider the dose.

Table 3 shows the evaluation results of the gastric mucosal damage inhibitory action of SC. It was revealed that an oral dose of 50 to 100 (mg extract powder / kg body weight) had an inhibitory effect of about 60%.

[0039]

[Table 2]

[0040]

[Table 3]

[0041]

EFFECTS OF THE INVENTION According to the present invention, since it is a composition derived from a natural plant, there is no fear of side effects even if it is taken for a long period of time, it is excellent in antioxidant action and nitric oxide production suppressive action, and has a wide range of efficacy as an anti-ulcer agent. It has become possible to provide a drug having

   ─────────────────────────────────────────────────── ─── Continued front page    (72) Inventor Sadashi Kageura             Rosebe, 1-5-13 Jinnan, Shibuya-ku, Tokyo             Lee Jinnan 3rd floor Nippon Bio Co., Ltd. (72) Inventor Yutana Kompiriyadecha             Kyoto, Kyoto City Yamashina-ku, Goryo 4 chome-cho 1 Kyoto             Inside Pharmaceutical University F-term (reference) 4B018 MD61 ME06 ME08 ME14 MF01                 4C088 AB12 AC01 BA07 BA08 NA14                       ZA66 ZA68 ZB01 ZC41

Claims (4)

[Claims] 1. An antioxidant containing one or more plants selected from the group consisting of Salacia quinensis, Salacia oblonga and Salacia reticulata or an extract thereof as an active ingredient. 2. A nitric oxide production inhibitor comprising, as an active ingredient, one or more plants selected from the group consisting of Salacia quinensis, Salacia oblonga and Salacia reticulata, or an extract therefrom. 3. An anti-ulcer agent containing, as an active ingredient, one or more plants selected from the group consisting of Salacia quinensis, Salacia oblonga and Salacia reticulata, or an extract therefrom. 4. A processed food containing one or more plant powders selected from the group consisting of Salacia quinensis, Salacia oblonga and Salacia reticulata or an extract thereof.