JP2014073992A - Helianthus tuberosus extract having pollinosis symptom reducing action - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a helianthus tuberosus extract which exhibits the production inhibiting action of an IgE antibody and a histamine, and has pollinosis symptom reducing action, and is useful as a food composition or a medicinal composition, and to provide a method for producing a helianthus tuberosus extract.SOLUTION: The present invention relates to the helianthus tuberosus extract hardly containing an inulin of a main ingredient or containing only an extremely little amount of the inulin, which is obtained by extraction using an extraction solvent with an inulin solubility of about 2% or less, for example, a mixture of one lower alcohol, which is selected from methanol, ethanol, propanol, and butanol, and water, more specifically, for example, hydrous alcohol with a water content of about 60-90%. The present invention also relates to the food composition or the medicinal composition which contains the extract as an active ingredient.

Description

The present invention relates to an extract of Jerusalem artichoke having an action of suppressing production of IgE antibody and histamine and exhibiting a remarkable symptom-reducing effect of hay fever, and a method for producing the same.

The present invention also relates to a food composition or a pharmaceutical composition containing the extract as an active ingredient.

Jerusalem artichoke (Helianthus tuberosus) is a perennial plant belonging to the genus Comositae sunflower, the main component of which is a dietary fiber containing the polysaccharide inulin, unlike ordinary potatoes, it contains almost no starch, Inulin has attracted attention as a low-calorie health food material because it has a blood glucose-lowering effect.

Up to now, as for the extract of Jerusalem artichoke, a multifunctional skin external preparation having a radical scavenging action, an elastase activity inhibitory action and a B16 cell whitening action, or a hair external preparation having an antioxidant action and a phospholipase A2 activity inhibitory action There is a report on a Jerusalem artichoke extract useful as an active ingredient (see Patent Document 1), or a Jerusalem artichoke extract useful as an active ingredient in a cosmetic composition for anti-inflammatory and regenerating skin (Patent Document 2).

Patent Document 1 describes an extract obtained by extracting Jerusalem artichoke with water or 50% ethanol or 50% 1,3-butylene glycol. This extract is mainly composed of inulin, the main component of Jerusalem artichoke, from the extraction solvent used. On the other hand, the Jerusalem artichoke extract of the present invention is obtained by extraction with an extraction solvent having an inulin solubility of about 2% or less, such as about 60-90% hydrous alcohol, and contains little or very little inulin. It is completely different from the extract.
In addition, in Patent Document 2, in addition to the extract obtained by extracting Jerusalem artichoke with water, the extract obtained by leaving Jerusalem for 4 days with 70% aqueous ethanol is filtered, and is cooled from light and air. Extracts stored protected are described. This extract overlaps with the extract of the present invention in terms of the extraction solvent. However, the Jerusalem artichoke extract of the present invention is obtained by filtering and purifying the extracted extract with an extraction solvent having an inulin solubility of about 2% or less, for example, about 60-90% hydrous alcohol, and concentrating and drying it. The Jerusalem artichoke extract is completely different in the composition and shape of the product.
Furthermore, the extract of the present invention exhibits an IgE antibody and histamine production-inhibiting action, and is completely different from the conventional extract in action and effect.
So far, no Jerusalem artichoke extract having an effect of suppressing the production of IgE antibody and histamine and showing a symptom-relieving effect of hay fever has not been reported or suggested.

Regarding the alleviation of symptoms of hay fever, such as sneezing and itching, it is said that it is effective to suppress the production of IgE antibody and histamine which are important inducers in hay fever.

As a result, the present inventor conducted research to find a plant extract component exhibiting an IgE antibody and histamine production inhibitory effect. As a result, the present inventor is an extract of Jerusalem artichoke that contains almost no inulin as a main component. Alternatively, the desired effect can be found in an extract containing very little, and the present invention has been completed.

JP2007-246446A Japanese Patent Publication No. 5-52805

An object of the present invention is to provide a Jerusalem artichoke extract having an effect of suppressing the production of IgE antibody and histamine and having an effect of reducing the symptoms of hay fever, and a food composition or a pharmaceutical composition containing the extract as an active ingredient. is there.

The inventors of the present invention have made extensive studies to solve the above-mentioned problems and have completed the present invention. That is, an extract of Jerusalem artichoke, which contains little or very little inulin as the main component, exhibits high IgE antibody and histamine production inhibitory action and has an effect of reducing the symptoms of pollinosis As a result, the present invention was completed.

It is known that inulin, which is the main component of Jerusalem artichoke, exhibits a hypoglycemic action, and several actions and effects of a Jerusalem artichoke extract mainly composed of inulin have been reported. (For example, see Patent Documents 1 and 2)
However, there is no report or suggestion about the component that exhibits the inhibitory action on IgE antibody and histamine production, which is an object of the present invention. The present inventors paid attention to components other than inulin, which is the main component of Jerusalem artichoke, and studied an extract containing little or very little inulin as the main component.

Inulin is known to have extremely high water solubility, while water-containing alcohol has a lower inulin solubility in proportion to the alcohol concentration. For example, it was confirmed that the inulin solubility of water-containing methanol or water-containing ethanol was extremely low at about 60% at a concentration of about 1.5 to 1.6%. From this, in order to obtain an extract that contains as little as possible the main component inulin, extract the Jerusalem artichoke using a hydroalcohol having an inulin solubility of about 2% or less, such as 70% methanol, and examine the extract. As a result, it was surprisingly found that the present invention showed remarkable IgE antibody and histamine production-suppressing effects and exerted symptom-reducing effects of hay fever. The present invention relates to a novel Jerusalem artichoke extract exhibiting such excellent effects.

More particularly, the present invention provides:
[1] An extract of Jerusalem artichoke (Helianthus tuberosus), which contains little or very little of the main component inulin,
[2] The extract according to [1], which exhibits an IgE antibody and histamine production-inhibiting action and has an effect of reducing symptoms of hay fever,
[3] The extract according to [1], which is obtained by extracting tuber of Helianthus tuberosus or a dry powder thereof using an extraction solvent having an inulin solubility of about 2% or less,
[4] The extract according to [3], wherein the extraction solvent is a mixture of one kind of lower alcohol selected from methanol, ethanol, propanol and butanol and water,
[5] The extract according to [4], wherein the extraction solvent is about 60 to 90% aqueous methanol or about 60 to 90% aqueous ethanol, or
[6] Jerusalem artichoke (Helianthus tuberosus) tuber or its dry powder is 1 to 100 times, about 60 to 90% aqueous methanol or about 60 to 90% aqueous ethanol, and a temperature condition of 4 to 60 degrees Celsius The extract according to [5], which is obtained by filtering and purifying an extract obtained by standing or stirring for 0.5 to 48 hours, and concentrating and drying the extract.
The present invention also provides:
[7] A method for producing a Jerusalem artichoke extract, characterized in that a tuber of Helianthus tuberosus or a dry powder thereof is extracted using an extraction solvent having an inulin solubility of about 2% or less,
[8] The method for producing a Jerusalem artichoke extract according to [7], wherein the extraction solvent is a mixture of one lower alcohol selected from methanol, ethanol, propanol and butanol and water, [9] Is a method for producing a Jerusalem artichoke extract according to [8], which is about 60-90% aqueous methanol or about 60-90% aqueous ethanol, and
[10] Jerusalem artichoke (Helianthus tuberosus) tuber or dry powder thereof is 1 to 100 times, about 60 to 90% aqueous methanol or about 60 to 90% aqueous ethanol, and a temperature condition of 4 to 60 degrees Celsius The method for producing a Jerusalem artichoke extract according to [9], wherein the extract obtained by standing or stirring for 0.5 to 48 hours is filtered and purified, and concentrated and dried. , Regarding.
Furthermore, the present invention provides
[11] A food composition or a pharmaceutical composition containing the Jerusalem artichoke extract according to [1] as an active ingredient,
[12] The food or pharmaceutical composition according to [11], which has an effect of reducing hay fever symptoms, and
[13] The present invention relates to a hay fever symptom alleviating agent comprising the Jerusalem artichoke extract according to [1] as an active ingredient.

The novel Jerusalem artichoke extract of the present invention has a remarkable IgE antibody and histamine production-inhibiting action, and is extremely useful as an active ingredient of a food composition or a pharmaceutical composition having an effect of reducing hay fever symptoms. is there.

FIG. 1 is a graph showing changes over time in the number of scratches in each of the Jerusalem artichoke extract group, normal group and control group obtained in Example 1. The vertical axis is the number of scratches (times) per 15 minutes, the horizontal axis is the number of intake days (days) until the measurement date, the value is the mean value ± standard deviation, and the “*” mark in the graph is the Student t- Each test indicates that there is a significant difference from the control at p <0.05. . FIG. 2 is a diagram showing the number of sneezes after 40 days of intake in each of the Jerusalem artichoke extract group, normal group and control group obtained in Example 1. The vertical axis is the number of sneezing (times) per 30 minutes, the value is the mean ± standard deviation. In the graph, “*” indicates a significant difference with respect to the control with p <0.05 by Student's t-test, respectively. It represents that there is. FIG. 3 is a graph showing the total IgE antibody titer in serum 40 days after ingestion of the Jerusalem artichoke extract group, normal group and control group obtained in Example 1. The vertical axis indicates the absorbance at 490 nm, the value is the mean value ± standard deviation, and the “*” mark in the graph indicates that there is a significant difference from the control at p <0.05 by Student's t-test. FIG. 4 is a graph showing Cry j 1-specific IgE antibody titers in serum 40 days after ingestion of each of the Jerusalem artichoke extract group, normal group and control group obtained in Example 1. The vertical axis indicates the absorbance at 490 nm, the value is the mean value ± standard deviation, and the “*” mark in the graph indicates that there is a significant difference from the control at p <0.05 by Student's t-test. FIG. 5 is a graph showing blood histamine concentrations 40 days after ingestion of the Jerusalem artichoke extract group, normal group and control group obtained in Example 1. The vertical axis indicates the absorbance at 490 nm, the value is the mean value ± standard deviation, and the “*” mark in the graph indicates that there is a significant difference from the control at p <0.05 by Student's t-test.

Hereinafter, embodiments of the present invention will be described in detail.
The present invention relates to an extract of Jerusalem artichoke, which contains little or very little of the main component inulin.
Jerusalem artichoke is a perennial plant belonging to the genus Sunflower, which is characterized by producing tubers, and its main component is dietary fiber containing the polysaccharide inulin, and unlike ordinary potatoes, it contains almost no starch. Moreover, since inulin exhibits a hypoglycemic action, it is attracting attention as a raw material for functional foods in a wide range of fields such as health maintenance.

The Jerusalem artichoke extract of the present invention is characterized in that it contains little or very little inulin as the main component.
Inulin, the main ingredient of Jerusalem artichoke, has extremely high water solubility, while water-containing alcohols have a lower solubility in proportion to the alcohol concentration.

In order to find an extraction solvent with low solubility of inulin, the present inventors conducted an inulin solubility measurement test of hydrous alcohol as follows.
Measuring method Ten times the amount of water-containing methanol or water-containing ethanol (concentration: 0 to 100%) was added to the crushed ground rice cake, and the mixture was stirred at room temperature for 2 hours to obtain an extract.
The extract was centrifuged (12,000 rpm, 15 minutes, 10 ° C.) to obtain a supernatant.
The amount of inulin contained in the extract and each supernatant before centrifugation was converted to the amount of fructose equivalent by the phenol-sulfuric acid method, and the solubility was calculated by the following equation.
Solubility = {(inulin amount of supernatant) / (inulin amount of solution before centrifugation)} × 100

The results of measurement are as shown in Table 1.

By this test, the inulin solubility of water-containing methanol or water-containing ethanol decreases to about 4-5% at a concentration of about 50%, rapidly decreases to about 1.5-1.6% at a concentration of about 60%, It was confirmed that about 90% was about 0.3 to 0.4%, which was almost the same as 100% alcohol.
The present invention is based on this finding, and that the Jerusalem artichoke tuber or its dry powder was extracted using an extraction solvent having an inulin solubility of about 2% or less, for example, about 60-90% hydrous alcohol. It is a feature.

Below, the outline of the manufacturing method of the Jerusalem artichoke extract of this invention is described.
That is, an extraction solvent having an inulin solubility of 1 to 100 times is added to Jerusalem artichoke tubers or dry powder thereof, and left at a temperature of 4 to 60 degrees (Celsius) for 0.5 to 48 hours. Alternatively, the crude extract is obtained by stirring. The purified extract obtained by subjecting the crude extract to filtration or centrifugation is subjected to operations such as vacuum concentration using an evaporator or spray drying, or spray drying to produce the Jerusalem artichoke extract of the present invention.

The extraction solvent may be any solvent that is acceptable as an extraction solvent for pharmaceuticals or food composition components and has an inulin solubility of about 2% or less, but methanol, ethanol, propanol, and butanol. A mixture of one kind of lower alcohol selected from the above and water is preferable, and a water-containing mixture of ethanol and methanol is more preferable among them.
The concentration of the hydrous alcohol may be any concentration as long as the solubility of inulin is about 2% or less. For example, in the case of hydrous methanol or hydrous ethanol, the concentration is within the range of about 60% to 90%. Although it can select suitably, about 70 to 80% is preferable.

The temperature condition is preferably about 4 to 60 ° C, and most preferably about 4 to 25 ° C.

In order to suppress the change of components as much as possible during extraction,
Is preferably adjusted to 4.0 to 9.0.

In the animal test using BALB / c mice, blood antibody titer and histamine concentration induced by sensitization of cedar pollen allergen Cry j 1 were measured for the Jerusalem artichoke extract of the present invention thus produced.

In other words, 5-week-old female BALB / c mice (Charles River) were divided into 3 groups, one of which was the normal group (no treatment group), and the other two groups were sensitized with the cedar pollen allergen Cry j1. Treatment was made into a control group and a Jerusalem artichoke extract group. Mice were sensitized by intraperitoneal injection of Cry j 1 (Nippon Biochemical Biobusiness) solution (15 μg / 150 μl) with Alum (Thermo) three times after 0, 7, and 14 days. Furthermore, nasal sensitization was performed by injecting the Cry j 1 solution (7 μg / 70 μl) into the nasal cavity of the mouse once a day for 14 days after 28 days. For normal and control groups, basic feed (MF feed, Oriental Yeast Co., Ltd.) is used, and for Kikuimo extract group, Kikuimo extract-added feed (basic feed containing 0.05% of Kikuimo extract) is used. Feeding was continued until day 40, and the number of scratches per 15 minutes was measured at 5-day intervals during the test period. Further, on the 40th day, the number of sneezes per 30 minutes was measured. Further, after completion of the test, blood was collected from the abdominal vena cava, and the total IgE antibody titer, Cry j 1-specific IgE antibody titer, and histamine concentration in the serum were measured by ELISA. In order to maintain the sensitization during the feed intake period, the Cry j 1 solution (1.5 μg / 15 μl) was injected into the nasal cavity of the mouse once a week.

As a result, after the 25th day, the number of scratches in the Jerusalem artichoke extract group was significantly reduced as compared with the control group. In addition, on the 40th day, the number of sneezing in the Jerusalem artichoke extract group was significantly reduced as compared with the control group.

Further, the total IgE antibody titer and Cry j 1-specific IgE antibody titer in the serum of the Jerusalem artichoke extract group were significantly lower than those in the control group. (P <0.05) The histamine concentration in the serum of the Jerusalem artichoke extract group was significantly lower than that in the control group (p <0.05).
This experimental result shows that the Jerusalem artichoke extract of the present invention has a remarkable IgE antibody and histamine production inhibitory action and exhibits an excellent hay fever symptom reducing effect.

Thus, the extract of the present invention exhibits an excellent hay fever symptom reducing effect, and can be used as an active ingredient in various food compositions or pharmaceutical compositions.

In the case of food, it can be used as various functional health foods in combination with appropriate food additives.
Moreover, when it is set as a pharmaceutical, it can be used as various dosage forms according to the method of a normal dispensing in combination with a suitable pharmaceutical additive. Examples of such dosage forms include oral preparations such as solid preparations such as powders, granules, capsules, pills and tablets, liquids such as liquids, suspensions and emulsions.

The dosage when the extract of the present invention is used as an active ingredient of a pharmaceutical composition varies depending on various factors such as the age, weight, sex, symptom level, and administration method of the patient. In the case of oral administration, it can be selected in the range of about 1-1000 mg, and in the case of parenteral administration, it can be selected within the range of about 0.1-100 mg. Moreover, it can also be appropriately increased or decreased depending on the degree of symptom improvement. The number of administration can be divided into once to several times a day.

The amount of intake when the extract of the present invention is used as a food can be selected according to the case of oral administration of the pharmaceutical product. However, in the case of food and drink, unlike pharmaceutical products, the dose and frequency of administration are not particularly limited, so the purpose of maintaining health, taste, and palatability should be considered unless particularly serious symptoms occur. And you may select intake, without being limited to said range.

Hereinafter, embodiments of the present invention will be described with reference to examples. However, the present invention is not limited to the following examples.

Manufacture of 70% methanol extract Rhizome of Jerusalem artichoke cultivated in Achi Village, Nagano Prefecture was collected. This was washed with water, dried with a dryer, and pulverized with a pulverizer to obtain a crushed potato.
After adding 300 mL of 70% methanol to 50 g of ground kikumo, the mixture was stirred and extracted overnight at 4 ° C. The supernatant obtained by centrifuging this (7500 × g, 10 minutes) was subjected to an evaporator to remove the solvent, and 16.1 g of Jerusalem artichoke extract was obtained.

Preparation of 70% ethanol extract According to the method of Example 1, using 70% ethanol as an extraction solvent, a Jerusalem artichoke extract was obtained as follows.
After adding 30 mL of 70% ethanol to 5 g of ground kikumo, the mixture was stirred and extracted overnight at 4 ° C. The supernatant obtained by centrifuging this (7500 × g, 10 minutes) was subjected to an evaporator to remove the solvent, and 2.89 g of a Jerusalem artichoke extract was obtained.

Japanese cedar pollen allergen Cry
j 1 Sensitization-induced blood antibody titer and histamine concentration measurement test About the Jerusalem artichoke extract obtained in Example 1, with respect to the blood antibody titer and histamine concentration induced by cedar pollen allergen Cry j 1 sensitization, The effect of oral intake was measured.
Five-week-old female BALB / c mice (Charles River) are divided into three groups, one of which is the normal group (no treatment group), and the remaining two groups are treated with cedar pollen allergen Cry j1 sensitization. The control group and the Jerusalem artichoke extract group were used.
Mice in the control group and the Jerusalem artichoke extract group were sensitized by intraperitoneal injection of Cry j 1 solution (15 μg / 150 μl) three times after 0, 7, and 14 days. Furthermore, nasal sensitization was performed by injecting the Cry j 1 solution (7 μg / 70 μl) into the nasal cavity of the mouse once a day for 14 days after 28 days.
In the normal group and the control group, the basic feed (MF feed, Oriental Yeast Co., Ltd.) was used, and in the Kikuimo extract group, the Kikuimo extract-added feed obtained in Example 1 (0.05% of the Kikuimo extract was added to the basic feed). %)) And continued until day 40. In order to maintain the sensitization during the feed intake period, the Cry j 1 solution (1.5 μg / 15 μl) was injected into the nasal cavity of the mouse once a week.
During the test period, the number of scratches per 15 minutes was measured at 5-day intervals. On the 40th day, the number of sneezes per 30 minutes was measured.
After completion of the test, blood was collected from the abdominal vena cava, and the total IgE antibody titer, Cry j 1-specific IgE antibody titer, and histamine concentration in the serum were measured by ELISA. For total IgE, add the above serum to an ELISA microplate coated with anti-mouse IgE antibody, bind IgE in the serum, react with horseradish peroxidase (HRP) labeled anti-rabbit IgG antibody, Color development by о-phenylenediamine (OPD) was measured with a microplate reader. The antibody titer was evaluated by absorbance at 490 nm (detection wavelength of the chromogenic substrate OPD of HRP, which is a label of the secondary antibody). For Cry j 1 specific IgE, biotinylated anti-mouse IgE monoclonal antibody and streptavidin labeled with horseradish peroxidase were used in the above protocol. About histamine, the density | concentration was measured by ELISA kit (Abnova). The histamine concentration was calculated from a calibration curve prepared by taking the absorbance of histamine of known concentration on the vertical axis and the concentration on the horizontal axis.

The test results are shown in FIGS.
As shown in FIG. 1, after the 25th day, the number of scratches in the Jerusalem artichoke extract group was significantly lower than that in the control group.
Moreover, as shown in FIG. 2, on the 40th day, the number of sneezes in the Jerusalem artichoke extract group was significantly reduced as compared with the control group.
As shown in FIG. 3, the total IgE antibody titer and Cry j 1-specific IgE antibody titer in the serum of the Jerusalem artichoke extract group were significantly lower than those in the control group. (P <0.05)
Moreover, as shown in FIG. 4, the histamine concentration in the serum of the Jerusalem artichoke extract group showed a significantly lower value compared with the control group (p <0.05).

The Jerusalem artichoke extract of the present invention has a remarkable IgE antibody and histamine production inhibitory action and has an effect of reducing the symptoms of hay fever. By containing this as an active ingredient, functional health food or hay fever Etc. can be manufactured.

Claims (13)

An extract of Helianthus tuberosus containing little or very little inulin as the main ingredient. The extract according to claim 1, which exhibits an effect of suppressing production of IgE antibody and histamine and has a symptom-reducing effect of hay fever. The extract according to claim 1, which is obtained by extracting tubers of Jerusalem artichoke (Helianthus tuberosus) or a dry powder thereof using an extraction solvent having an inulin solubility of about 2% or less. The extract according to claim 3, wherein the extraction solvent is a mixture of one lower alcohol selected from methanol, ethanol, propanol and butanol and water. The extract according to claim 4, wherein the extraction solvent is about 60-90% aqueous methanol or about 60-90% aqueous ethanol. Jerusalem artichoke (Helianthus tuberosus) tuber or dry powder thereof is 1 to 100 times, about 60 to 90% aqueous methanol or about 60 to 90% aqueous ethanol at a temperature of 4 to 60 degrees Celsius, and 0 The extract according to claim 5, which is obtained by filtering and purifying an extract obtained by standing or stirring for 5 to 48 hours, and concentrating and drying the extract.   A method for producing a Jerusalem artichoke extract, comprising extracting a tuber of Helianthus tuberosus or a dry powder thereof using an extraction solvent having an inulin solubility of about 2% or less.   The method for producing a Jerusalem artichoke extract according to claim 7, wherein the extraction solvent is a mixture of one lower alcohol selected from methanol, ethanol, propanol and butanol and water. The method for producing a Jerusalem artichoke extract according to claim 8, wherein the extraction solvent is about 60 to 90% aqueous methanol or about 60 to 90% aqueous ethanol. Jerusalem artichoke (Helianthus tuberosus) tuber or dry powder thereof is 1 to 100 times, about 60 to 90% aqueous methanol or about 60 to 90% aqueous ethanol at a temperature of 4 to 60 degrees Celsius, and 0 The method for producing a Jerusalem artichoke extract according to claim 9, wherein the extract obtained by standing or stirring for 5 to 48 hours is filtered and purified, and then concentrated and dried.   A food composition or a pharmaceutical composition comprising the Jerusalem artichoke extract according to claim 1 as an active ingredient.   The food composition or pharmaceutical composition according to claim 11, which has an effect of reducing hay fever symptoms. A hay fever symptom alleviating agent comprising the Jerusalem artichoke extract according to claim 1 as an active ingredient.

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