KR101394524B1 - Anti-histamine substance and method for production thereof - Google Patents

Abstract

A method for producing an antihistamine material according to the present invention comprises: a step of adding yeast and water to a dried stevia plant tissue to effect fermentation;

Extracting fermented stevia obtained by the fermentation process using an aqueous ethanol solution;

Concentrating the extract obtained by the extracting step; And

And collecting the 90% ethanol-soluble fraction by fractionating the concentrate obtained by the concentration step.

Description

[0001] Anti-histamine substance and method for production thereof [

The present invention relates to an allergic symptom-improving substance which causes hay fever, urticaria, bronchial asthma, atopy and the like, and a method for producing the same.

Allergy refers to an immune response that occurs excessively for a particular antigen. Allergies are largely classified into type I to V according to their mechanism of occurrence, and pollen, chlamydia, and bronchial asthma are classified as type I. It is also considered to be close to allergy to type I atopy. It is thought that type I allergy occurs because histamine or leukotrienes released from these cells by IgE antibody binding to mast cells or basophils are released to the whole body.

It is known that the active ingredient is a fermented concentrate of Stevia stigma (Staphylococcus epidermidis) as an internal medicine for treating gastrointestinal diseases, which is effective for treating external skin diseases such as atopic dermatitis or for treating gastritis and gastric ulcer. Further, it is known that fermented plant tissue extract of Stevia is known as a substance having an antihistaminic effect that is effective for allergic symptoms such as hay fever.

The inventors of the present invention have also found that, in view of the problem that the stevia plant tissue fermentation takes longer than 90 days in the above-described technology, the present inventor has proposed a method of reducing the time required for fermentation of Stevia extract, The stevia extract is pulverized and mixed, and the mixture is placed in purified water and heated, stirred, cooled and filtered to obtain a filtrate. The filtrate is concentrated under reduced pressure to obtain stevia X. The stevia extract is then fermented by adding yeast bacteria (Japanese Patent Application Laid-Open No. 2005-35888).

As described above, many improvement substances such as hay fever and atopy are already proposed and put on the market. In order to improve the degree of improvement of the allergic symptoms more than now, it is common for many sufferers. Among these situations, research and development have been actively pursued in search of an allergy symptom improving substance superior to the conventional one.

It is an object of the present invention to provide a substance exhibiting superior antihistamine action, in particular, in order to help improve the symptoms of type I allergy.

After repeated research to solve the above problems, the present inventors have succeeded in producing a substance having antihistamine action far superior to that of the prior art, using Stevia plant tissue as a raw material. The method for producing an antihistamine material according to the present invention having the above-mentioned effect comprises the steps of adding fermented Stevia plant tissue obtained by the fermentation step with yeast and water to the dried stevia plant tissue, extracting using the aqueous ethanol solution, A step of concentrating the extract obtained by the extracting step and a step of collecting the 90% ethanol-soluble fraction by fractionating the concentrate obtained by the concentrating step.

The inventors of the present invention have found that a strong antihistaminic action is recognized in a substance obtained through the above-mentioned production method. Therefore, this substance can be widely applied to cosmetics suitable for a person who shows symptoms of type I allergies, external preparations, internal replication.

In addition, the material obtained by the above-described production method has no unpleasant odor. On the other hand, the antihistamine material obtained by the production method described in Patent Document 1 has an unpleasant odor. Since the substance obtained by the present invention has no such problem, there is an advantage that it is easier to use than the conventional substance when it is incorporated into an external preparation, particularly a cosmetic. Therefore, the present invention includes cosmetics and medicines (external preparation, internal replication) containing the substance obtained by the above-mentioned production method in the range.

1 is a flow chart showing a flow of a method for producing an antihistamine material according to the present invention.

FIG. 2A is a graph showing one of the experimental results of the antihistamine effect of the substance of the present invention. FIG.

FIG. 2B is a view showing one of the experimental results of the antihistamine effect of the substance of the present invention.

FIG. 2C is a graph showing one of the results of an experiment for examining the antihistamine effect of the substance of the present invention. FIG.

FIG. 3 is a graph showing one of the experimental results of antihistamine effect of the substance of the present invention. FIG.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings.

First, a method for producing the antihistamine material according to the present invention will be described in detail with reference to the flowchart of FIG.

Step 1 is a step of drying and pulverizing the plant tissue of Stevia as a raw material. One of the raw materials of the present invention, Stevia plant is a perennial plant of Chrysanthemum which is originated from South American Paraguay, and its scientific name is Stevia Rebaudiana Bertoni (Stevia Rebaudiana Bertoni). When cultivated in Japan, it is often the case that the seedlings, seedlings or the seedlings are planted and planted from February to May. Harvesting is often done from October through November, when stevia plants are mature enough.

The antihistamine material according to the present invention uses dry Stevia powder prepared by drying the plant tissue of the stevia well and grinding it finely. It is sufficient to crush the powder so that the size of each piece is approximately 1 cm or less. It is also preferable to selectively use leaves and stem parts of Stevia plants.

Drying of leaves and stems of Stevia plants is preferably carried out at room temperature in a well-ventilated room as possible, and it is not desirable to apply abrupt heat.

Step 2 is a step of fermenting the dry Stevia powder obtained in Step 1. As described above, the method for producing an antihistamine material according to the present invention is characterized in that dry Stevia powder is directly fermented. On the other hand, in the manufacturing method disclosed in Japanese Patent Application Laid-Open No. 2005-35888, the extract is first extracted from the dried Stevia powder, and the extracted extract is fermented. However, according to the results of subsequent studies conducted by the inventors of the present invention, it has been found that a substance that is strongly fermented with dry Stevia powder can produce a substance having strong antihistamine effect.

The fermentation of the dried Stevia powder can be carried out by adding water and yeast to the dry Stevia powder, stirring and leaving. The amount of water to be added may be as much as the amount necessary for fermentation, and the amount enough to wet the whole is sufficient. As the yeast, it is preferable to use Saccharomyces.

It is preferred that the fermentation of the dried Stevia powder is performed " completely ". If you taste the unfermented stevia powder, it will be sweet, but you will not feel the sweetness in the fully fermented stevia powder. Therefore, it is preferable to proceed the fermentation until no sweet taste is felt at all. In order to fully ferment, approximately 2 to 3 weeks of incubation period is required at room temperature.

As a kind of yeast, Saccharomyces species are used. In the fermentation process, moisture is first given to dried Stevia powder, the yeast is added thereto, and the fermentation is continued while supplying water for 2 to 3 weeks at room temperature. The determination of whether or not the fermentation of the stevia powder is "complete" can be made, for example, by considering the time when the sweetness is not felt by putting the stevia powder in the mouth as the "complete" fermentation end.

Step 3 is a step of extracting and concentrating the fermented Stevia powder obtained in Step 2, and it is important to use ethanol for the extraction. This is because it is one of the characteristics of the present invention that the 90% ethanol-soluble fraction is collected in the next step 4.

The ethanol extraction can be carried out by preparing a 30% aqueous ethanol solution, adding the fermented Stevia powder thereto, and allowing the mixture to slowly infiltrate at room temperature for several hours to several days. The filtrate is filtered with a size of about 140 mesh, and the resulting filtrate is used as an extract. In the production method according to the present invention, the extract is concentrated by reducing the pressure at a temperature of about 60 캜.

Step 4 is a step of collecting the 90% ethanol-soluble fraction by fractionating the concentrated extract obtained in Step 3. The fraction can be carried out by passing the concentrated extract through an ion exchange resin column such as DIAION (registered trademark) HP-20. A strong antihistamine action is recognized in the fractionated ethanol soluble fraction.

The 90% ethanol-soluble fraction fractioned for convenient storage and transportation may be frozen and dried (Step 5). This completes the process for producing the antihistamine material in the preferred embodiment of the present invention.

<Test Example>

Pharmacological experiments on antihistaminic action were carried out on the substance of the present invention obtained through steps 1 to 5 shown in Fig. 1, using a specimen of a guinea pig ileum (ileum). The experiment was carried out by extracting the ileum from the guinea pigs and measuring the isometric contraction induced by histamine by the Magnus method.

10 ^-6 M histamine to Marmot (histamine10 ^-6 M) when applied to the sample extraction president is induced contraction. Therefore, histamine 10 ^-6 M was applied before and after application of the substance of the present invention to examine the effect of the substance of the present invention on the contractile reaction by observing the contraction reaction difference. The shrinkage was recorded with a DC amplifier (AM20, manufactured by Unipulse Corporation) through a tension transducer (BG-10, Kulite Semiconductor, USA). Histamine was obtained from Wako Pure Chemical Industries, Ltd.

For the application of the substance of the present invention, a solution was prepared by dissolving the substance of the present invention in a nutrient solution (Lock-Ringer solution). Three kinds of test solutions having different concentrations of the substance of the present invention were prepared, and the concentrations of the substance of the present invention were adjusted to 10 ^-5 g / ml, 10 ^-4 g / ml, and 10 ^-3 g / ml, respectively. The solution was applied directly to the margins tube.

The composition of the rocking solution was NaCl = 154, KCl = 5.6, CaCl ₂ = 2.2, MgaCl ₂ = 2.1, NaHCO ₃ = 5.9, and glucose = 2.8 (pH = 7.4). The nutrient solution was a mixture of 95% O ₂ and 5% CO ₂ .

Experiments were carried out at room temperature (23-24 ° C).

The experimental results are shown in Figs. 2A to 2C. Figure 2a, the material density of the present invention ¹⁰ is the experimental results for the ⁵ g / ㎖ of solution. At 6:22, 10 ^-6 moles of histamine was applied to the specimen of the guinea pig excised site to determine the contraction reaction, which was called the control group. After the contraction of the specimen was completed, a solution of the inventive substance concentration 10 ^-5 g / ml was applied to the specimen at 6:30, and histamine 10 ^-6 mol was again applied to the specimen at 6:33. In this measurement, the recording graph by AM20 manufactured by UNIFULS Corporation was as shown in Fig.

The horizontal axis of the graph of FIG. 2A represents time, and represents one minute of two minutes. The vertical axis represents the voltage and is approximately proportional to the contraction size of the venous specimen (Figures 2b and 2c are the same). When the shrinkage size at the first application of histamine was 100, the shrinkage size at the second application of histamine was 89.7 on the graph. The decrease in shrinkage size during the second application of histamine is thought to be due to the antihistamine effect of the solution.

FIG. 2B is a recording graph of AM20 produced by UNIPHERM Co., Ltd. as a result of a test on a test solution having a concentration of the present invention of 10 ^-4 g / mL. At 4:47, 10 ^-6 moles of histamine was applied to the specimens of the guinea pig excised site to determine the contraction reaction, which was called the control group. After the contraction of the specimen was completed, the test solution of the present invention at a concentration of 10 ^-4 g / ml was applied to the specimen at 4:55, and histamine 10 ^-6 mol was again applied to the specimen at 4:58.

When the concentration of the present invention substance in the test solution is 10 ^-4 g / ml, the shrinkage size at the time of the second application of histamine for the first time when the shrinkage size at the time of histamine application (the voltage value of the DC amplifier) is 100 is 60 . Therefore, it can be seen that a strong antihistamine effect is exerted as compared with the case where the substance concentration of the present invention is 10 ^-5 g / ml.

2C is a recording graph showing the results of experiments on a test solution of the present invention concentration of 10 ^-3 g / ml. At 6:40, 10 ^-6 moles of histamine was applied to a specimen of the guinea pig excised site to determine the contraction reaction, which is referred to as the control group. After the contraction of the specimen was completed, a solution of the inventive substance concentration of 10 ^-3 g / ml was applied to the specimen at 6:50, and histamine 10 ^-6 mol was again applied to the specimen at 6:52.

As can be seen from Fig. 2C, when the concentration of the present invention substance is 10 &lt; ^-3 &gt; g / ml, no shrinkage reaction of the specimen occurs at the second application of histamine. This was an unexpected and surprising result even for the present inventors. Taking this result into consideration, it can be understood that the antihistaminic effect of the substance of the present invention is strong beyond conventional common sense.

FIG. 3 is a graph summarizing the experimental results of FIGS. 2 (a) to 2 (c), wherein the abscissa represents the logarithm of the concentration of the substance of the present invention in the test solution and the ordinate represents the ratio of the shrinkage response measured after the application of the test solution and the control. As the concentration of the present invention substance in the solution increases, it is easy to understand that the shrinkage reaction is greatly reduced.

The details of the method for producing the antihistamine material according to the present invention and the experimental results for illuminating the effect of the antihistamine material produced by the method are shown above. From the experimental results, it has been shown that the antihistamine substance according to the present invention has a remarkable antihistamine effect. Therefore, the antihistamine material according to the present invention can be widely applied to cosmetics and pharmaceuticals, and can be expected to be applied to cosmetics or pharmaceuticals suitable for people with type I allergies, such as hay fever or atopy. Furthermore, the antihistamine material according to the present invention is characterized by no unpleasant odor, and this makes it possible to easily apply to cosmetics sensitive to odor problems. Therefore, the antihistamine material according to the present invention is expected to be very important as a component to be incorporated in cosmetics for people having symptoms such as hay fever or atopy.

The antihistamine material according to the present invention is useful as a cosmetic for hair cosmetics, a hair lotion, a hair lotion, a hair dye, a hair coloring agent, a hair lotion, a hair rinse, a skin cosmetics / lotion, a cosmetic liquid, a cream, a milky lotion, Cosmetics, foundation, makeup base, min, lipstick, eye make-up, ball cosmetics, body make-up, odecoron, perfume, cosmetics, body lotion, facial rinse, pack, cosmetic oil, body rinse, , Cosmetics for baths, nail cosmetics, body powders, etc. In addition, medicines include powders, granules, tablets, capsules, pills, sticks, pencils, , Aerosols, gas agents, drug adsorbents, ophthalmic solutions, injections, bandages and the like.

Claims (6)

As a method for producing an antihistamine substance, A fermentation process in which the dried Stevia plant tissue is directly fermented by adding yeast and water to the dried stevia plant tissue, An extraction step of extracting the fermented stevia obtained by the fermentation step using an aqueous ethanol solution, A concentration step of concentrating the extract obtained by the extraction step, and And a step of subjecting the concentrate obtained by the concentration step to ethanol fractionation. The method according to claim 1, Wherein the stevia plant tissue is completely fermented in the fermentation step. Extracting the fermented Stevia obtained by directly fermenting the dry Stevia plant tissue by adding yeast and water to the dried stevia plant tissue and allowing it to stand, extracting the obtained stevia with an ethanol aqueous solution, concentrating the extract and ethanol fractioning the resulting concentrate Antihistamine substance. A cosmetic comprising the antihistamine substance produced by the production method according to claim 1 or 2, or the antihistamine substance according to claim 3. An antihistamine substance produced by the method according to claim 1 or 2, or an antihistamine substance according to claim 3. delete

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