JP2008007473A - Cassia obtusifolia l. or cassia tora l. seed extract - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a Cassia obtusifolia L. or Cassia tora L. seed extract which contains a clinically effective amount of a rubrofusarin glycoside and reduces the content of a specific ingredient to an extent not inducing diarrhea. <P>SOLUTION: The Cassia obtusifolia L. or Cassia tora L. seed extract contains (A) emodin, aloe-emodin, chrysophanol, physcion and aurantio-obtusin, and their glycosides, and (B) rubrofusarin and its glycoside in a (A)/(B) weight ratio of ≤2.00 (preferably ≤1.90, more preferably ≤1.80, further preferably ≤1.75), obtained by controlled extraction. The controlled extraction is performed by finely crushing the Cassia obtusifolia L. or Cassia tora L. seeds and then immersing the finely crushed product in hot water. The finely crushing treatment is preferably performed by an explosion treatment or a germination treatment. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

The present invention relates to the provision of a kameiko extract containing a high concentration of rubrofusarin glycoside and a low concentration of diarrhea-inducing component.

Conventional technology

Ketsumeishi is a seed of Ebisu rush, roasted and roasted, drunk as hub tea, and used as a folk medicine because it is effective for eye symptoms. In Japan, Seiko is listed as a herbal medicine in the pharmacopoeia and is decocted for the purpose of treating the bowel, abdominal bloating, and constipation. There is also an attempt to find an antioxidative action in the Seiko extract and use it as an antioxidant for cosmetics containing fats and oils (Patent Document 1).

  As is clear from the fact that it is used for the purpose of intestinal regulation and improvement of bowel movement, the deciduous action has been recognized in the extract of Natsumeko (Non-patent Documents 1 and 2), and as an ingredient having such action Anthraquinone chrysophanol, emodin, aloe-emodin, rhein, physcion, obtusin, obtusifolin, chryso-obtusin, aurantio-obtuin and the like (Non-patent Document 3).

In recent years, it has been found that a component that promotes intracellular glutathione synthesis ability is present in the extract of Ketsumeishi, and it has been specified that this component is a rubrofusarin glycoside (Patent Document 2). Glutathione is a multifunctional biological component that is contained in cells at a relatively high concentration. In many diseases caused by oxidative stress and aging associated with aging, the intracellular glutathione level decreases. It recognized. Therefore, increasing the in vivo glutathione concentration is considered significant for the prevention of these diseases or conditions. However, even if glutathione itself is administered orally, its half-life in blood is as short as several minutes, so it is not effective for increasing glutathione levels in vivo.
Japanese Patent Laid-Open No. 10-8049 Japanese Patent Application Laid-Open No. 2004-2231 Michio Takido, “Resources and Ingredients of Seiko and Their Biological Activity”, Contemporary Oriental Medicine Vol. Chikako Tanaka and Ryuichi Kato, “NEW Pharmacology”, Nanedo (1992), p440 Shibata Seuni, “Natural Medicinal Substances”, Nanzando (1982), p457-470

  According to the study by the present inventors, an increase in blood glutathione concentration was observed when about 30 mg of rubrofusarin glycoside per day was orally administered to humans (see Reference Example 1 and FIG. 2).

  Further, according to the study by the present inventors, the ethanol extract of Akemiko contained a lot of rubrofusarin glycosides, but also had a laxative action. On the other hand, the hot water extract has low sputum action but low rubrofusarin glycoside content, and in order to consume an effective amount of rubrofusarin glycoside, a large amount of extract is consumed, It has been found that not only is it unfavorable, but the possibility of exerting an armpit action is increased.

  Since it is considered that the Keriko extract contains various active ingredients, there are various significances in taking a rubrofusarin glycoside as a Keriko extract, not just ingesting only a rubrofusarin glycoside. It can be said. However, as described above, ingestion of the Kakeko extract containing an effective amount of rubrofusarin glycoside for increasing the blood concentration of glutathione is accompanied by ingestion of a relatively large amount of components having a sputum action, such as diarrhea. There was concern that undesirable effects would be induced.

  From such a point of view, the present invention has an object to provide a Kameiko extract containing a clinically effective amount of rubrofusarin glycoside and having a specific component content reduced to such an extent that diarrhea is not induced. To do.

  According to the study by the present inventors, by performing controlled extraction under special conditions, the content of the diarrhea-inducing component and rubrofusarin glycoside in the Keriko extract was able to be set to a specific weight ratio. And the Kakeko extract thus obtained has been found to have no substantial depressive action even when used for the purpose of ingesting a clinically effective amount of rubrofusarin glycoside, thereby completing the present invention. .

That is, the present invention contains the following (A) and (B) in a weight ratio [(A) / (B)] of 2.00 or less (preferably 1.7 or less, more preferably 1.6 or less, and even more preferably 1.5 or less). Provide Keikoko extract:
(A) emodin, aloe emodin, chrysophanol, fission and aurantioobtusin, and their glycosides;
(B) Rubrofusarin and its glycoside.

In the present specification, the term “Keiko” refers to the seed of the Japanese name Ebisugusa ( Cassia obtusifolia L. or Cassiatora L.) of the leguminosae ( Leguminosae ), unless otherwise specified . The seeds may be used as they are or may be roasted, but in the present invention, those that are not roasted can be suitably used. The present invention can also be applied to a plant containing a rubrofusarin glycoside and a diarrhea-inducing component in the same manner, instead of Akemi. For example, Cassao occidentalis belonging to the genus Kawara-Ketsumei can also be used in the present invention in the same manner as Keiko.

  The group (A) of the present invention comprises components that can induce diarrhea contained in the Seiko extract. These include the anthraquinones, emodin, aloe-emodin, chrysophanol, physcion, obtusin, chryso-obtusin, orlanthioo. Examples include butausin-obtuin, obtusifolin, and rhein (see Non-Patent Document 3). Each of these components usually exists as a glycoside in the clarifier. In the present invention, as the diarrhea-inducing component group (A), emodin, aloe-emodin, chrysophanol, fission and aurantioobtusin can be used as indices. In the present specification, the term “emodin, aloe emodin, chrysophanol, fission and auranthioobtusin, and their glycosides” refers to emodin, aloe emodin, chestnut, unless otherwise specified. Consists of all existing sofanols, fissions, auranthioobtusins, emodin glycosides, aloeemodin glycosides, chrysophanol glycosides, fission glycosides and auranthioobtusin glycosides Ingredient group.

  The group (B) of the present invention consists of rubrofusarin and rubrofusarin glycoside. When all rubrofusarin exists as a glycoside, group (B) consists of only rubrofusarin glycoside. Rubrufusaline is usually present as a glycoside in Seiko and its extracts.

  The term “rubrofusarin glycoside” as used herein refers to rubrofusarin gentiobiosite and its analogs represented by the following formula, except for special cases. Rubrofusarin trisaccharide glycosides and tetrasaccharide glycosides also exist, and these are referred to herein as rubrofusarin glycosides.

  In this specification, the weight of (A) when referred to as “weight ratio [(A) / (B)]” is emodin, aloe emodin, chrysophanol, fission and aurantioobtusin except in special cases. Refers to the total weight. That is, those that are not glycosides are left as they are, and those that are glycosides are the emodin, aloeemodin, chrysophanol, fission and auranthioobtusin excluding the glycoside moiety, and the total weight of them. Point to. In the present specification, the weight of (B) when referred to as “weight ratio [(A) / (B)]” refers to the weight as rubrofusarin. That is, what exists as rubrofusarin remains as it is, and rubrofusarin glycoside refers to the total weight of rubrofusarin excluding the glycoside moiety. Those skilled in the art can appropriately determine the weight of (A) and the weight of (B). For example, in accordance with the method shown in Example 1 of the present specification, the glycoside moiety was removed by treating the Seiko extract with a hydrolase (eg, β-glucosidase), and the glycoside moiety was removed by HPLC. By quantifying the target component, the weight of (A) and the weight of (B) can be determined. In the present invention, the weight ratio [(A) / (B)] is 2.00 or less, preferably 1.90 or less, more preferably 1.80 or less, and still more preferably 1.75 or less.

Preferred form for carrying out the invention

  In order to make the weight ratio [(A) / (B)] as described above, the clarifier is finely pulverized; the finely pulverized product is 60 ° C. or higher (preferably 70 ° C. or higher, more preferably 80 ° C. or higher, More preferably, the extract may be obtained by immersing in hot water at 90 ° C. or higher for 35 to 300 minutes (preferably 40 to 240 minutes, more preferably 150 minutes, and further preferably 45 to 120 minutes).

  The seed pulverization method includes a treatment for softening the seed tissue such as a germination treatment, an HHS treatment (high-temperature high-pressure treatment), an explosion treatment (instantaneous high-temperature pressure treatment), and a pH treatment. Explosive treatment or germination treatment is preferred.

The blasting treatment refers to a process in which a raw material is pulverized by exposing it to high temperature and high pressure conditions for a predetermined time in a pressure vessel and then rapidly releasing it to atmospheric pressure. In the present invention, there are no particular limitations on the conditions for blasting the clarifier as long as the desired weight ratio can be achieved. For example, 158 to 225 ° C. (saturated vapor pressure 5 to 25 kg / cm) used in ordinary blasting equipment. ² G), the conditions of seconds to hours can be applied. When applying the conditions, it may be considered that the main component, rubrofusarin glycoside, is relatively resistant to heat. By subjecting the water to vaporization and explosion, the volume of the clarifier expands explosively by subjecting it to the blasting treatment, so that the structure of the clarifier can be sufficiently crushed from the inside, and the desired components from the inside of the tissue can be removed in the subsequent extraction process. Extraction efficiency can be increased.

  In the present invention, the germination treatment can be carried out by placing the deciduous seeds in a sufficiently water-containing state for 1 to several days under heating as necessary. Germination can be confirmed visually, and when a large number of seeds are treated, the time when germination of a part (for example, one) of them can be confirmed can be used as a reference for termination of germination treatment. Germination treatment can also increase the extraction efficiency of the desired components from within the tissue of Komeiko seed.

  As a method for fine pulverization of Akemiko seeds in the present invention, the normal mechanical pulverization process has a finer raw material particle size, so the load in the subsequent extraction step is large, the yield decreases, and becomes inefficient. It may not be preferable. Further, in the pulverization process, since the structure of Akemi is usually not broken, the extraction efficiency of a desired component is inferior compared with the blasting process and the germination process.

  The finely pulverized product of Keiko is used for the extraction process. The extraction solvent is not particularly limited as long as the desired [(A) / (B)] weight ratio can be achieved, and examples thereof include an aqueous solvent (for example, a mixture of water, water and ethanol), and ethanol. In ethanol extraction, a lot of diarrhea-inducing components are extracted, and in water, extraction of diarrhea-inducing components can be relatively controlled. Therefore, an aqueous solvent, particularly water, is preferable.

  As long as the desired [(A) / (B)] weight ratio can be achieved, an organic solvent may be used, and two or more organic solvents may be used in combination. From the viewpoint of production, an aqueous or ethanol solvent, particularly water, is preferred.

  The extraction temperature can be made relatively high because the main component, rubrofusarin glycoside, is resistant to heat. For example, when water is used as the solvent, the temperature is 60 ° C. or higher, preferably 70 ° C. or higher, more preferably 80 ° C. or higher, and still more preferably 90 ° C. or higher. A pressure device is required to set the temperature to 100 ° C. or higher, and the temperature may be set to 100 ° C. or lower in consideration of the reduced amount of preparation.

  The extraction time is, for example, 35 to 300 minutes, preferably 40 to 240 minutes, more preferably 45 to 120 minutes. According to the study by the present inventors, when hot water at 90 ° C. was used, there was no difference between the extraction for 60 minutes and the extraction for 120 minutes. Considering this result, it may be 60 minutes or less.

  The extract obtained by the extraction step may be further subjected to enzyme treatment and column operation. Enzyme treatment is a treatment for removing viscous substances (polysaccharides) produced after hot water extraction to increase production efficiency. The enzymes used are commercially available α-amylase, β-amylase, cellulase, Pectinase and the like can be used, and α-amylase is particularly preferable. As the column operation resin, an aromatic synthetic adsorbent can be suitably used.

  The following is an example of preparing the extract of the present invention using Seiko as a raw material. Solvent is added to the blasted or germinated Koriko, heated and extracted, filtered for solid-liquid separation, and the filtrate is treated with enzyme to obtain a crude extract filtrate. The amount of solvent added for extraction can be 3 to 10 times the amount of the raw material. The filtrate can be column processed. For column operation, for example, an adsorption resin such as Diaion HP20 (manufactured by Mitsubishi Chemical Corporation) that is widely used in industry is used. The hot water extract can contact the filtrate directly with the resin. Fractionate by adsorbing impurities on the column. The extract thus obtained can be concentrated or dried into powder by a conventional method such as concentration under reduced pressure or freeze-drying.

  This invention also provides the food-drinks containing the Keiko extract obtained by this invention. Such foods and drinks should be designed to contain an amount of rubrofusarin glycoside effective to maintain or increase glutathione concentration in the blood, since the deciduous action of the Keriko extract of the present invention is reduced. Can do.

  In the present specification, “food and beverage” includes seasonings, nutritional supplements, health foods, dietary foods, general health foods, supplements and beverages, and orally administered medicines.

  The specific shape of the food or drink may be any shape. Examples thereof include solid, semi-fluid, and fluid. Examples of solid foods include general foods and health foods in the form of biscuits, sheets, tablets such as tablets and capsules, and granular powders. Examples of semi-fluid foods include pastes, jellies, and gels. Examples of fluid foods include general foods and health foods in the form of juices, soft drinks, teas, and drinks. The food / beverage products of the present invention are preferably in the form of tablets, beverages and the like.

  The blending amount of Keriko extract in the food and drink of the present invention is not particularly limited, but referring to the preferred daily intake of rubrofusarin glycoside, those skilled in the art, depending on the intake form of the food and drink, A compounding quantity can be set suitably. Usually, it adds to the food / beverage products of this invention so that it may contain 30 mg or more, 45 mg or more, 60 mg or more of rubrofusarin glycosides per daily intake.

  The food / beverage products of the present invention can contain various components used in ordinary food / beverage products, in addition to the Keriko extract of the present invention. Examples of these additives and / or ingredients include vitamins, sugars, excipients, disintegrants, binders, lubricants, emulsifiers, tonicity agents (isotonic agents), buffers, solubilizers, Examples include antiseptics, stabilizers, antioxidants, colorants, flavoring agents, fragrances, coagulants, pH adjusters, thickeners, tea extracts, herbal extracts, inorganic salts, and the like.

  In addition, on the food and drink of the present invention, its specific use (for example, for health maintenance etc.) and / or its specific usage (for example, intake, number of intakes, intake method, etc.) shall be displayed. Can do.

The present invention also provides (A) a weight ratio of (E) emodin, aloe emodin, chrysophanol, fission and auranthioobtusin, and their glycosides to (B) rubrofusarin and its glycosides [(A) / (B)] Maintaining or increasing the glutathione concentration containing the Kakucho extract, characterized by being reduced to 2.00 or less (preferably 1.90 or less, more preferably 1.80 or less, and even more preferably 1.75 or less) The oral composition for reducing the swallowing action; and pulverizing the clarifier; the pulverized product at 60 ° C. or higher (preferably 70 ° C. or higher, more preferably 80 ° C. or higher, more preferably 90 ° C. or higher). Glutathione concentration containing Kameiko extract, which includes the step of immersing in hot water for 35 to 300 minutes (preferably 40 to 240 minutes, more preferably 150 minutes, more preferably 45 to 120 minutes) to obtain an extract. Maintenance or oral composition for increasing, provides a method of reducing the cathartic effect.
〔The invention's effect〕

  The present invention uses as an index the active ingredient rubrofusarin glycoside that increases the ability to synthesize glutathione in cells and the identified diarrhea inducing component, and the content weight ratio of rubrofusarin glycoside to the diarrhea inducing component [(A) / (B)] The present invention provides a kameiko extract that can efficiently ingest active ingredients and has a low risk of inducing diarrhea as a composition that stipulates that it is 2.00 or less.

  EXAMPLES Hereinafter, although an Example demonstrates this invention, this invention is not limited only to an Example.

[Example 1] Analysis of content ratio of rubrofusarin glycoside and diarrhea-inducing component under various extraction conditions
1. Materials and methods
Seiko Extract (A) Seiko (Chinese, C.tora dried seed) was extracted with 70% (v / v) ethanol at 50 ° C for 2 hours, filtered through 60 mesh, Diaion HP20 ( A powder (brown powder) obtained by spray drying a solution operated by Mitsubishi Chemical Corporation.

  (I) Seiko (Chinese, C.tora dried seed) extracted with 10 times the amount of hot water at 90 ° C for 1 hour, filtered through 60 mesh, and subjected to enzyme treatment with α-amylase at 60 ° C for 15 minutes. A powder (brown powder) obtained by spray drying a solution operated with Diaion HP20 (Mitsubishi Chemical Corporation).

  (C) Seiko (China, C. tora dry seed) was blasted at 170 ° C for 1.2 seconds, extracted with 20 times the amount of hot water at 90 ° C for 1 hour, filtered through 60 mesh, and α-amylase Powder (brown powder) obtained by spray-drying a solution treated with Diaion HP20 (manufactured by Mitsubishi Chemical Corporation) at 60 ° C for 15 minutes.

Analytical sample preparation method
The enzyme β-glucosidase (manufactured by Yakult Co., Ltd.) was dissolved in 0.6 M acetate buffer (pH 4.9) at 1 mg / ml, and an extract previously dissolved in ethanol was added to a concentration of 1 mg / ml. And reacted for 1.5 hours. The same amount of 0.1% TFA / CH ₃ CN as the reaction solution was added as a reaction stop solution, and after sonication, a solution filtered through a 0.45 μm filter was prepared as an analysis sample. This was subjected to HPLC, and the amount of each component was determined based on a calibration curve prepared in advance using a standard substance.

HPLC analysis condition column: Ascentis C18-3μm, 4.6mmφ x 150mm
Mobile phase: A = 0.1% TFA / H ₂ O, B = 0.1% TFA / 100% CH ₃ CN, 0.8ml / min
Gradient: B40% → B80% (10min), B80% iso10min
Injection: 10μL, detection: A280nm (PDA: 220-500nm)
2. The results of confirming the dissolution pattern are shown in Fig. 1. The analysis results are summarized in Table 1 (in the table, the unit of each component amount is μg / mg).

  From Table 1, the weight ratio of (a) ethanol extraction, (b) hot water extraction, (c) blasting hot water extraction each of ruburofusarin and diarrhea-inducing components is about twice that of (c) Yes, (I) is about 1.3 times (U). In other words, (a) and (b) are extracted about twice as much and about 1.3 times as many diarrhea-inducing ingredients as compared to the active ingredient rubrofusarin. .

[Example 2] Analysis of the content ratio of rubrofusarin glycosides and diarrhea-inducing components by Setsuko Germination
Extraction method of germinated Keriko Keriko (produced in India) germinated in 3 days, extracted with hot water at 90 ° C, treated with α-amylase, and operated with Diaion HP20 (Mitsubishi Chemical) The extract was used.

  Preparation and analysis were performed in the same manner as in Example 1.

  The results are shown in Table 2.

  From Table 2, the content weight ratio of rubrofusarin and diarrhea-inducing component was 1.55. This can be said to be an extraction method that can reduce the diarrhea-inducing properties equivalent to or higher than (c) explosion-treated hot water extraction in Table 1.

[Example 3] Diarrhea-inducing test of Akemiko extract
ICR male mice 6 weeks old (CLEA Japan, Inc.), using 10 to 11 mice per group, the extract was orally administered twice at a rate of 10 ml / kg at intervals of 5 hours, and activated carbon 30 minutes later Was further administered 30 minutes later, and the charcoal powder transport rate (%) in the small intestine [movement distance of charcoal powder (cm) × 100 / total length of the small intestine (cm)] was measured.

  The results are shown in Table 3.

  From Table 3, it is clear that (a) ethanol extract and (c) hot water extract and (c) explosive-treated hot water extract show a diarrhea-induced reduction. In particular, the explosion-treated hot water extract of (c) can reduce diarrhea-inducing properties and has a large amount of extracted rubrofusarin glycoside.

[Example 4] Preparation of beverage containing Keriko extract Extract 0.9g Keriko purified according to the extraction method of Example 1 (c) was dissolved in 189.1g of pure water and sterilized by retort (121 ° C, 15 minutes). A beverage was prepared. Table 4 shows.

  It was light brown and did not smell, and the taste was faint and bitter. The rubrofusarin content was about 8 mg / tube.

[Example 5] Production of food containing Kameiko extract 90 g of Kameiko purified extract produced according to the extraction method of (c) of Example 1 is mixed, and a tablet is produced using a tableting machine. Table 5 shows.

[Example 6] Preparation of Kakeyoko extract-containing food Tea leaves such as cereal tea, oolong tea, green tea and black tea are used alone or mixed to obtain 5 to 20% of the total amount of the final product to obtain an extract. To the extract, Kakusei refined extract produced according to Example 2 (final concentration of about 0.4%), ascorbic acid (final concentration of about 0.4%), and sodium bicarbonate (final concentration of about 0.25%) were added to a pH of about 6.0. Prepare to ˜7.0 and add pure water to make a tea drink.

[Reference Example 1] Effect of intake of KAMEKO on glutathione concentration in human plasma 5 beverages containing KAMEKO ethanol extract (content of KAMEKO extract: 0.3 g / 190 mL) in 5 healthy subjects with average bowel movements Cans / day (35.6 mg / day as rubrofusarin) × 3 days, then 3 cans / day (106.7 mg / day as rubrofusarin) × 3 days were taken, and the results of 4 persons were obtained.

  The measurement results of plasma glutathione (GSH) concentration for four people are shown in FIG. As a result of the interview, most people answered that bowel movements became loose.

  According to the present invention, for the purpose of preventing or treating anti-fatigue, it becomes possible to provide beverages and foods, in particular, health foods, with Keriko extract, which can be safely consumed with less side effects of diarrhea.

It is a HPLC analysis chart which shows a composition of Kakeyoko extract. It is a graph showing the raise of the glutathione (GSH) density | concentration in the human plasma by Kaseki child intake. The left graph shows the average value of four people, and the right graph shows the change of each value.

Claims (8)

Seiko extract containing the following (A) and (B) at a weight ratio [(A) / (B)] of 2.00 or less:
(A) emodin, aloe emodin, chrysophanol, fission and aurantioobtusin, and their glycosides;
(B) Rubrofusarin and its glycoside.   A clarified extract obtained by an extraction method comprising a step of pulverizing clarified powder; dipping the pulverized material in hot water at 60 ° C. or higher for 35 to 300 minutes to obtain an extract.   Food-drinks containing the extract of Claim 1 or 2.   The food or drink according to claim 3, comprising an amount of rubrofusarin glycoside effective for maintaining or increasing the glutathione concentration in blood. The manufacturing method including the step of controlling and extracting the following (A) and (B) so that the weight ratio [(A) / (B)] is 2.00 or less:
(A) emodin, aloe emodin, chrysophanol, fission and aurantioobtusin, and their glycosides;
(B) Rubrofusarin and its glycoside.   6. The production method according to claim 5, wherein the controlled extraction step is a step of finely pulverizing the clarifier; and immersing the finely pulverized product in hot water at 60 [deg.] C. or higher for 35 to 300 minutes to obtain an extract. Glutathione concentration in vivo containing Kakucho extract, characterized by reducing the following (A) to a weight ratio [(A) / (B)] 2.00 or less with respect to (B) below A method for reducing the swallowing effect of an oral composition for maintaining or increasing
(A) emodin, aloe emodin, chrysophanol, fission and aurantioobtusin, and their glycosides;
(B) Rubrofusarin and its glycoside.   An oral composition for maintaining or increasing the glutathione concentration containing the clarifier extract, comprising a step of pulverizing the clarifier; immersing the finely pulverized product in hot water at 60 ° C. or higher for 35 to 300 minutes to obtain an extract. Of reducing the armpit action.

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