JPH11217333A - Antiallergic agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an antiallergic agent having weak toxicity and adverse effect and exhibiting excellent antiallergic effect by making the agent include plant body itself of a plant belonging to the genus Rhodiola and containing an antiallergic substance in the plant body or its extract. SOLUTION: This antiallergic agent contains plant body itself (roof, rhizome, leaf, etc., or a cultured cell obtained by subjecting the root, rhizome, etc., to callus culture according to ordinary method) of a plant belonging to the genus Rhodiola and containing an antiallergic substance in its plant body (e.g. Rhodiola sachalinesis or Rhodiola ishidae) or its extract (e.g. an extract extracted from a plant body with an aqueous solvent or a water-soluble organic solvent) as an active ingredient. The effective daily dose as dried material of ethanol extract is usually about 0.1-2 g/kg weight in the case of oral administration and about 10-250 mg/kg weight in the case of parenteral administration.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to an antiallergic agent. The antiallergic agent of the present invention is used for foods such as antiallergic foods, quasi-drugs such as cosmetics and creams for atopic dermatitis, pharmaceuticals and additives for compound feeds for preventing allergic diarrhea and the like. It is possible.

[0002]

2. Description of the Related Art At present, as means for preventing or treating allergies, steroid hormones and many nonsteroidal synthetic drugs have been developed in the pharmaceutical field. In addition, in foods and feeds, research is progressing in the development of removing allergens that cause allergies, that is, in the direction of de-allergenization.

[0003] In pharmaceuticals, there are problems such as side effects due to long-term administration of the drug.
It is impossible to deallergenize any food or feed material, and even if it is developed, it has drawbacks such as a problem in taste and an increase in product cost.

[0004] In addition, the antiallergic and antiinflammatory effects of unsaturated fatty acids such as EPA and DHA contained in fish oil are known, and it has been reported that their ingestion can change the allergic constitution. It needs to be taken on a daily basis, and there has been a demand for an appropriate combination with a component having an effect in a short term and its material.

[0005]

SUMMARY OF THE INVENTION Accordingly, an object of the present invention is to provide an antiallergic agent which has low toxicity and side effects and exhibits an excellent antiallergic effect.

[0006]

Means for Solving the Problems As a result of earnest studies, the present inventors have found that a plant of the genus Rhodiola contains an antiallergic substance exhibiting an excellent antiallergic effect in a plant, and completed the present invention. did. That is, the present invention provides an anti-allergic agent containing a plant itself or an extract thereof as an active ingredient, which belongs to the genus Rhodiola and contains an anti-allergic substance in the plant.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION As a preferred example of a Rhodiola plant used in the present invention, Rhodiola sacarinensis ( Rh
odiola sachalinensis ) and Rhodiola isidae ( Rhod)
iola ishidae ) and the like, but are not limited thereto. Rhodiola sacarinensis is known as Kojingen, a dicotyledonous plant known as Crassulacea.
It is a long-standing herbaceous plant belonging to e). The vegetation of Hongjing Heaven is
It is a mountainous area from 1800m to 2300m above sea level, and is mainly distributed in Asia (China, Japan, North Korea), Europe and North America.

[0008] In the present invention, the root of the plant of the genus Rhodiola,
Any part such as rhizome, stem and leaf can be used, but preferably root or rhizome is used. In addition, cultured cells obtained by callus-culturing cells such as roots or rhizomes by a conventional method can also be used.

The antiallergic agent of the present invention may be a plant such as roots, rhizomes, stems and leaves, or cultured cells, or may be an extract extracted therefrom. Antiallergic substances contained in Rhodiola plants are
It can be extracted from plants using an aqueous solvent or a water-soluble organic solvent. These solvents include water, lower aliphatic alcohols (methanol, ethanol, propanol)
And mixtures thereof. Also, an aqueous solution of an acid or an alkali can be used. Of these, water, ethanol and mixtures thereof are particularly preferred. In addition, when performing extraction with water or a low-concentration aqueous ethanol solution, 80 ° C.
Extraction can be performed more efficiently when heating is performed at about the boiling point. In the case of ethanol or a high-concentration aqueous ethanol solution (ethanol concentration of 60% or more), extraction can be performed efficiently even at room temperature, and extraction can be performed at any temperature from room temperature to the boiling point. The extraction time is appropriately selected depending on the solvent used and the extraction temperature, but is usually about 3 to 48 hours.

When the plant itself is used, a dried or freeze-dried plant is preferably crushed because it is easily added to feed or various compositions. The extract is preferably added to the product as it is, or dehydrated and concentrated by a known drying method such as a reduced-pressure drying method or a freeze-drying method, and is preferably used as a dried product because it is easily added to feeds and various compositions. At the time of drying, an excipient such as dextrin can be added to improve the shape of the dried product, and the production can be performed by a known process in consideration of the form of use.

The antiallergic agent of the present invention exerts its effects in both oral and parenteral administration. Parenteral administration includes topical administration such as application to the skin, intravenous injection,
Examples include subcutaneous injection, intramuscular injection, enteral administration, eye drops, nasal administration, intraperitoneal administration and the like.

The effective dose of the antiallergic agent of the present invention is as follows:
Although it is appropriately selected according to the symptoms and intended use, it is usually about 0.1 g to 2 g in the case of oral administration of 1 kg body weight per day as a dry product of the ethanol extract, and in the case of parenteral administration, Is about 10 mg to 250 mg.

When used as a pharmaceutical, the antiallergic agent of the present invention can be mixed with other excipients and formulated into a desired form according to a conventional method. Further, it can be added to quasi-drugs such as cosmetics and creams for atopic dermatitis and the like, and can be added to foods and compound feeds. The concentration when added to cosmetics, creams and the like is not particularly limited, but is preferably about 10 mg to 50 mg per 1 g of the product composition as a dry product of the ethanol extract. In addition, when added to food or compound feed, as an ethanol extract dried product, usually 2 kg per 1 kg of food or feed is used.
g to about 20 g are preferred.

The antiallergic agent of the present invention suppresses the release of leukotriene (LTB ₄ ) and histamine from rat mast cells upon antigen stimulation and an inflammatory reaction involved in prostaglandin, as specifically shown in the following Examples. Has a remarkable suppressing effect on Further, as specifically described in Examples below, the antiallergic agent of the present invention has no acute toxicity when administered orally, and has a very low acute toxicity even when administered intraperitoneally. Also, unlike steroids and the like, it is a natural product, so its safety is high.

[0015]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below more specifically based on embodiments. However, the present invention is not limited to the following examples.

Example 1 Preparation of Dried Extract Extract 100 g of a powdered red rhizome of the red rhizome was
After adding 6 times volume of 70% ethanol, extracting at room temperature for 24 hours and filtering the extract, adding 4 times volume of 70% ethanol to the residue and extracting for another 24 hours, filtering the extract and further extracting the residue. The operation was repeated to perform three extraction operations. The extracted filtrates were mixed three times, concentrated under reduced pressure, and dried at 70 ° C. under reduced pressure to obtain 15 g of a dried product. The dried extract was used for anti-allergy and anti-inflammatory tests.

Example 2 Preparation of Dried Extract Extract To 6 g of 99.5% ethanol was added to 100 g of the same dry powder of red Kageten as in Example 1, and the mixture was shaken at 50 ° C. for 1 hour.
After extracting for 2 hours, filtering the extract, the same operation is performed on the residue, and the extract filtrate is mixed twice, and concentrated under reduced pressure.
Lyophilization yielded 16.5 g of a dried product.

Example 3 Solubility of Extracted Dry Product 0.5 g of each of the extracted dried products obtained in Example 1 was dissolved in 10 times the amount of various solvents, and the solubility was tested. The results are shown in Table 1 below. In addition, p when dissolved in 10 times the amount of water
H was 4.2.

[0019]

[Table 1]

Example 4 Antiallergic Effect As one of means for confirming the antiallergic effect, there is a method of observing the effect of suppressing the release of leukotriene (LTB ₄ ) and histamine from rat mast cells. Therefore, the following test was conducted to confirm the antiallergic effect of the dried and dried red Kagekeiten product obtained in Example 1.

That is, a 10% phosphate buffer solution (PBS) of the dried and dried red seaweed extract obtained in Example 1 was heated at 121 ° C.
After heating in an autoclave for 15 minutes, the mixture was centrifuged, and the supernatant was used as a sample. Rats are 7 week old Splague Dawr
Using y male rats, intraperitoneal cells including mast cells (hereinafter, “PEC”) were obtained from the peritoneal cavity, and the inhibitory effect of the extracted sample on the release of LTB ₄ and histamine by stimulation of Ca ionophore was examined. Was. This test was specifically performed as follows.

That is, a rat was sacrificed by ether anesthesia, 30 ml of Tyrode's solution was injected into the abdominal cavity, and the abdomen was removed.
After massaging for a minute, the abdomen was opened and the Tyrode solution was collected.
The collected solution was centrifuged at 200 g for 5 minutes, and the supernatant was removed. The number of cells was adjusted with a Tyrode solution according to the following experiment and used for the following experiment.

The measurement of LTB ₄ was performed using 854 μl of Tyrode solution,
Red Kageten extract (10% PBS extract) 10μl, 50μ
100 μl of M-Ca ionophore solution, 25 mM-CaC
a mixture of l ₂ solution 36μl were prepared as a reaction solution.

2 × 10 ⁶ cells / ml prepared in advance
Was centrifuged at 200 g for 5 minutes, and after removing the supernatant, 50 μl of the reaction solution was added, followed by culturing at 37 ° C. for 20 minutes.
After cultivation, acetonitrile / methanol mixture (6: 5)
The reaction was stopped by adding 50 μl. After stopping the reaction, 12,0
00 were centrifuged 10 minutes at rpm to obtain a supernatant, it was subjected to determination of LTB ₄ by HPLC.

Histamine is 10 ⁶ cells / m in Tyrode solution.
1 ml of PEC prepared in 1 liter,
BS extract) 250 μl, 50 μM-Ca ionophore 250 μl, 25 mM CaCl ₂ solution 90 μl, and Ty
After mixing 910 μl of the rode solution and culturing at 37 ° C. for 20 minutes,
The reaction was stopped by cooling on ice. After stopping the reaction, 4 ° C, 200
g, and centrifuged for 5 minutes, and the supernatant was subjected to quantification of histamine by fluorescence analysis.

The results are shown in Table 2 below. As shown in Table 2, L from PEC treated with the extracted sample of Hongjing Heaven
Release of TB ₄ and histamine control (saline)
Less than 2% in the LTB ₄ as compared to, the histamine approximately 1
0% or less, the PE of these allergic substances
It was confirmed that release from C was significantly suppressed.

[0027]

[Table 2] *: Significantly different from control (P <0.05)

[0028] Example 5 in experimental system using an anti-inflammatory effect rat mast cells, LTB ₄ and implementation is possible to suppress the release from mast cells of histamine Example 4 dry extract of Benikeiten to elicit an allergic reaction Was recognized. Therefore, the following test was performed to confirm whether or not the experimental inflammatory response caused by an allergen was suppressed.

That is, a physiological saline solution containing 10% of the dried and dried extract of Kokukeiten obtained in Example 1 was heated at 121 ° C. and 15 ° C.
The mixture was autoclaved for minutes, and used as a sample solution. The rats were 6-week-old SD male rats, and 0.5 ml to 1 ml of the sample solution was administered intraperitoneally, allowed to stand for 30 minutes, and the thickness of the instep of the right hind leg was measured with a dial gauge. . Then, 0.1 ml of physiological saline containing 1% λ-carrageenin
Was injected subcutaneously into the right hind leg of the rat to induce edema and λ
-2, 3, 4, 5 hours after carrageenin inoculation, the thickness of the instep of the right hind foot was measured with a dial gauge. The inflammation rate was calculated from the thickness of the instep of the right hind foot after inoculation of λ-carrageenin,
Subtract the thickness before inoculation and subtract the value for saline inoculation by 10
The inflammation rate of the sample solution was calculated as 0.

The results are shown in Table 3 below. As shown in Table 3, the ethanol extract of Hongjingtian showed a remarkable effect on the experimental inflammatory reaction caused by λ-carrageenin. This study showed the same effect as suppressing action to carrageenan paw edema indomethacin prostaglandin biosynthesis inhibitors, anti-inflammatory potential of Benikeiten with LTB ₄ release inhibition effect from mast cells of Example 4 Proof.

[0031]

[Table 3]

Example 6 Acute toxicity (intraperitoneal administration) [0032] The acute toxicity of intraperitoneal administration of the extract of the dried red Kagekeiten obtained in Example 1 was examined. That is, the average weight is 23.7.
9 g of ICR mouse males (32 days old) were tested with 6 mice at each concentration level shown in Table 4 below, and inoculated with the solution of the dried extract of Kokukeiten extract into the peritoneal cavity of the mice. Observations were made for days, and mortality and survival rates were calculated. The LD ₅₀ calculated by the probit method was 3810 mg / kg body weight, which was extremely low toxicity.

[0033]

[Table 4]

Example 7 Acute toxicity (oral administration) [0034] The acute toxicity of the ethanol-extracted dried product of Kokukeiten obtained in Example 1 in oral administration was examined. That is, a sample was prepared from 10 male (5 week-old) ICR mice having an average body weight of 28.31 g so as to have a weight ratio of 5000 mg / kg body weight. Gavage was performed using a sonde, and the number of deaths and clinical findings after the administration were observed for 7 days. As a result, all the animals survived at one dose of 5000 mg / kg body weight, did not show any death or clinical abnormality, and were nontoxic.

[0035]

As described above, according to the present invention, an anti-allergic agent having low toxicity and excellent anti-allergic and anti-inflammatory effects was provided.

 ──────────────────────────────────────────────────続 き Continuing from the front page (72) Inventor Yasuo Samegai 1789-3 Kamishizu, Sakura-shi, Chiba Harmony Shizu B201 (72) Inventor Atsushi Hamano 1-67-67 Takamihara, Kamizakicho, Inashiki-gun, Ibaraki-pref. Inventor Fujiharu Tanji 3-7-16 Kakinokizaka, Meguro-ku, Tokyo

Claims (3)

[Claims] 1. An anti-allergic agent comprising, as an active ingredient, a plant itself or an extract thereof, which belongs to the genus Rhodiola and contains an anti-allergic substance in the plant. 2. The antiallergic agent according to claim 1, wherein the active ingredient is obtained by extracting the plant with water, ethanol, or a mixture thereof. 3. The antiallergic agent according to claim 1, wherein the plant is Rhodiola sacarinensis.