JPH11130685A - Antiallergic agent - Google Patents

Abstract

(57) [Summary] [Objective] To provide a highly safe, naturally occurring antiallergic agent. An antiallergic agent comprising, as active ingredients, an extract extracted from betel leaves using water or an organic solvent, and demethyleugenol isolated and purified from the extract.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to an allergic reaction,
In particular, the present invention relates to a natural product-derived antiallergic agent which is classified as type I allergy and is effective for so-called atopic allergy.

[0002]

2. Description of the Related Art Antiallergic drugs are drugs intended to prevent, suppress and alleviate allergic reactions. That is, the antiallergic agent is thought to act as a cell stabilizing factor and / or as a mast cell inhibitory factor that prevents mast cells such as histamine and leukotriene from degranulation that occurs in response to the presence of an antigenic substance. .
When an antigenic substance invades, degranulation occurs by binding to immunoglobulin IgE on the surface of mast cells, and mast cell components are excreted into the extracellular environment. Induces symptoms due to various allergic reactions.

As antiallergic agents, various synthetic drugs have been developed and used in addition to corticosteroids. However, their side effects are often always a problem. Further, among various allergic diseases, development of a drug for treating type I allergy, a so-called atopic symptom, whose onset has been significantly increased in recent years, is strongly desired.

[0004]

[0006] If there is a natural product that controls the onset of these allergic reactions by taking it easily in a tea-like form every day, the benefits will be enormous. In view of the above problems, the present inventors have sought natural plants used as foods and spices, not only in Japan, but also widely in China and Southeast Asia, using a model of type I allergy,
investigated.

[0005] In the course of the study, the present inventors focused on betel (English name: Piper betel). Betel is generally marketed as a crude drug in Southeast Asia and the like, but it is not known at all that betel itself and its extract exhibit an antiallergic effect.

[0006] Betel is an evergreen ivy shrub belonging to the pepper family, and is widely distributed in Southeast Asia, India, Sri Lanka and the like. Betel leaves, roots and seeds are used as a stomach and expectorant, especially from Taiwan to Southeast Asia, India,
In Sri Lanka and Pakistan, so-called betel
It is used as a chewing gum (chewing gum) and is used as a luxury item by chewing it like a gum, and is known as a sacred and important crude drug in Ayurvedic Indian Studies (traditional medicine in Sri Lanka, India).

[0007] In view of the above problems, the present inventor has continued systematic experimental studies in order to provide a novel antiallergic agent using a naturally derived crude drug with high safety, and as a result, completed the present invention. .

[0008] In other words, the fact that demethyleugenol, which is contained in betel leaves at a very high content (weight ratio: 1 to 3%), has an effect of suppressing an allergic reaction, particularly to type I which is an atopic allergic reaction The present inventors have found that they exhibit a strong inhibitory action, and have completed the present invention. It is completely unknown that betel leaves contain demethyleugenol and that demethyleugenol has an antiallergic effect.

[0009]

The present invention provides an antiallergic agent comprising as an active ingredient an extract extracted from betel leaves using water or an organic solvent, and an extract obtained by extracting from betel leaves using water or an organic solvent. The present invention relates to an antiallergic agent comprising, as an active ingredient, demethyleugenol represented by the chemical structural formula [2] obtained by isolating and purifying an extract. The chemical structural formula of demethyleugenol is shown below.

[0010]

Embedded image

[0011]

According to the present invention, in short,
Efficacy of the present invention can be confirmed by evaluating an allergy model using an extract obtained by extracting betel leaf powder with water or an organic solvent, or using demethyleugenol isolated and purified from the extract. Was. As the organic solvent, alcohol, acetone, or the like is used.

[0012]

BEST MODE FOR CARRYING OUT THE INVENTION The betel in the present invention uses a leaf portion which has been edible since ancient times. Estimated from the data of experimental animals described below, the dose per human is 1000-2000 mg for betel leaf extract, 15-30 mg for demethyleugenol, and sufficient anti-allergic
Show the effect. The dosage form used is granules or tablets with the addition of suitable excipients, but the effect of betel leaf powder as it is in the form of a tea preparation does not change.

Hereinafter, the present invention will be described with reference to examples and the like, but the present invention is not limited to the following examples.

Extraction method Extract 1 liter of solvent was added to 100 g of betel leaves 100-200 mesh obtained with a crusher, heated for about 3 hours, filtered, and the filtrate was concentrated under reduced pressure at 45 ° C. Then, the solvent was completely distilled off, and water, alcohol and acetone were used as solvents to obtain a water extract, an alcohol extract and an acetone extract, respectively. The yield of the water-extracted extract was about 22 to 23%, and the yield of other organic solvent-extracted extracts was about 15 to 23%.

Isolation and Purification of Demethyleugenol 2 kg of betel leaves , roughly cut and dried, were put into 10 liters of 35% alcohol, extracted with warming for about 3 hours, filtered, and the filtrate was completely decompressed at 45 ° C. under reduced pressure. To 35%
An alcohol extract was obtained. The extract yield was 23.2
%Met. The extract is applied to a silica gel column and n-
Hexane: ethyl acetate = 3: 1 eluted with a developing solvent,
About 24.1 g of demethyleugenol was obtained (yield: 5.
2%). That is, as shown in FIG. 1, the content of demethyleugenol in dried betel leaves was as high as 1.2%. Demethyleugenol is a compound already known in the literature, and it has been determined from various physical constants such as NMR that it is the compound.

Example 1 Passive cutaneous rat anaphylaxis (PCA) reaction of rat dinitrophenylated bovine serum albumin (hereinafter DNP)
-BSA) was prepared according to the method of Tada and Okumura (literature name: J. Immunol, 106, pp. 1002-1011 [1971]). Anti-DNP used was manufactured by Seikagaku Corporation. Tranilast (manufactured by Kissei Pharmaceutical Co., Ltd.) was used as a control. The experimental method used was a Wistar male rat (body weight 20).
0-220 g) of the back, and anti-D diluted 6250 times with phosphate buffered saline (pH 7.4) in the skin.
0.1 ml of NP was injected and sensitized. After 46 hours, each test drug was suspended in 1% gum arabic and orally administered. After 2 hours, 0.75 mg of DNP-BS
0.5 ml of A dissolved in 1% Evans blue was administered via the tail vein to induce a PCA reaction. Thirty minutes later, the blood was exsanguinated from the carotid artery and the area of the dye leaked to the back was measured using a digital planimeter (manufactured by Uchida Yoko Co., Ltd.). Was determined. Table 1 shows the results. The betel leaf extract, especially a 35% alcohol extract, had a strong antiallergic effect, and demethyleugenol was more effective than tranilast.

[0017]

[Table 1]

Example 2 Histami due to antigen stimulation from rat-sensitized peritoneal exudate cells
Tranilast (manufactured by Kissei Pharmaceutical Co., Ltd.) and Amlexanox (manufactured by Takeda Pharmaceutical Co., Ltd.) were used as free control drugs. The experiment was performed using Wistar male rats (body weight 30).
0-400 g) of a buffer solution for mast cells (pH 7.
4, hereinafter abbreviated as MCM), and gently massaged for 2 minutes. After laparotomy, the exuding liquid was collected with a pipette, and the peritoneal cavity was further washed with 10 ml of MCM to collect the exuding liquid. This is centrifuged (100 ×
g4 ° C., 10 minutes), the cells were suspended in 3 ml of MCM, and incubated with 1 ml of anti-DNP diluted 100-fold with MCM at 37 ° C. for 1 hour to perform passive sensitization. After completion of the incubation, the cells were washed three times with MCM, and then the number of mast cells was calculated using a hemocytometer under toluidine blue staining, and the cell suspension was finally adjusted to 10 ⁴ cells / ml. 1.62 ml of cell suspension at 37 ° C
After 10 minutes of incubation with each test drug solution 18
0 μl was added and reacted for 15 minutes. Then 200μl
Phosphatidylserine (1mg / ml) and 220μl
Of DNP-BSA (1 mg / ml) was added simultaneously and incubation was continued for another 20 minutes. The reaction was stopped by ice-cooling the test tube, and centrifugation (100 × g, 4 ° C., 10 minutes)
Thereafter, the histamine released by separating the supernatant was quantified by a fluorescence method. Table 2 shows the results. The betel leaf extract and its active ingredient demethyleugenol were found to suppress the release of histamine which causes inflammation and pain from mast cells.

[0021]

[Table 2]

[0022]

The antiallergic agent of the present invention is a natural product which is effective for various atopic symptoms including hay fever, and is based on the idea of pursuing a crude drug which has been edible since ancient times and which can be confirmed to have high safety. As a result of testing a large number of edible plants using an experimental animal model, betel leaves used mainly in Southeast Asia and the like showed excellent antiallergic effects equivalent to or higher than those of commercially available medical drugs, especially for atopic symptoms. It is also excellent in that it is a material that is effective for type I allergies and that can be supplied safely and at low cost. Therefore, it can be said that the industrial applicability of the present invention is very high.

[Brief description of the drawings]

FIG. 1. Isolation of demethyleugenol from betel leaves
It is a flow sheet showing a purification step and the like.

Claims (2)

[Claims] 1. An antiallergic agent comprising, as an active ingredient, an extract extracted from betel leaves using water or an organic solvent. 2. An antiallergic agent comprising, as an active ingredient, demethyleugenol represented by the chemical structural formula [1] obtained by isolating and extracting an extract obtained by extracting from betel leaves with water or an organic solvent. . Embedded image