JP2000327576A - MEDICAMENT FOR ANTI-SENSITIVE DISEASE CONTAINING 4G-alpha-D- GLUCOPYRANOSYLRUTIN - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject medicament having high water solubility, substantially tasteless and odorless, having high safety, capable of being easily produced, and useful for the prophylactic or therapy of sensitive diseases such as viral diseases, bacterial diseases, traumatopathy, by including a specific glucopyranosylrutin. SOLUTION: This medicament is obtained by including 4G-α-D- glucopyranosylrutin of the formula. The compound of the formula is obtained by acting a glycosyltransferase such as cyclomaltodextrin glucanotransferase and glucoamylase on a solution containing rutin and a starchy substance such as dextrin, normally at pH 3-10 and at 10-90 deg.C. It is desirable that the above- mentioned solution contains about 1.0-20.0 W/V% of the above-mentioned rutin, that the concentration of the above-mentioned starchy substance is at about 0.5-50 times that of the above-mentioned rutin, and that daily dose of this medicament is normally 0.001-50.0 g/adult calculated as the compound of the formula.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an anti-sensitive disease agent containing a novel substance ^4G- α-D-glucopyranosylrutin represented by the following chemical formula 1.

[0002]

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[0003]

2. Description of the Related Art Rutin has a chemical structure represented by the following chemical formula 2, and has long been known as a vitamin P having a physiological action such as strengthening of capillaries, prevention of bleeding, blood pressure regulation, and a yellow pigment. It is used for food, medicine, cosmetics, etc.

[0004]

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[0005] Vitamin P participates in vivo in the physiological activity of vitamin C, for example, the hydroxylation reaction of proline and lysine necessary for the synthesis of collagen, which is a main component of biological connective tissue. Fe
⁺⁺⁺ by reducing participate in redox reactions such as the Fe ^++, further, is known to enhance the activity of such involved in immune potentiation by leukocytosis, health maintenance of a living body, to enhance Plays an important role.

[0006] The use of rutin is not limited to vitamin P fortifier as a nutrient, but from its chemical structure and physiological action, alone or in combination with other vitamins, for example, yellow colorant, antioxidant, etc. As food and drink,
In addition, the range is extremely wide ranging from prophylactic and therapeutic agents for susceptible diseases such as cardiovascular diseases, that is, anti-susceptible agents, and even cosmetics as skin colorants such as yellow colorants and ultraviolet absorbers, and whitening agents. .

However, rutin is poorly soluble in water, and only about 1 g (about 0.01 W / V) is added to 8 L of water at room temperature.
%), Which is extremely difficult to use.

As a method for improving this, as disclosed in Japanese Patent Publication No. 25-1677, for example, a method of increasing the water solubility by adding an aliphatic compound having an amino group to rutin, and Japanese Patent Publication No. 26-2724. As disclosed in Japanese Patent Application Publication No. JP-A-2002-163, a method of reacting rutin with monohalogenacetic acid to form a sodium acetate compound to increase water solubility.
As shown in JP-A-1285, there has been known a method in which Rongalit acts on rutin to form a sulfite compound to increase water solubility.

However, in these methods, amino compounds, monohalogenacetic acid, sulfite compounds and the like are used, and the resulting substances are concerned with other physiological activities and toxicity.
Also, its purification is difficult.

Therefore, the present inventors have previously described Japanese Patent Publication No.
JP-A-32073, as a method of water-solubilization utilizing a biosynthesis reaction with higher safety, a glucose residue is transferred from a partial hydrolyzate of starch to rutin by an action of glycosyltransferase in an equimolar or more, and α- A method was proposed to produce glycosyl rutin and improve its water solubility.

As described in Japanese Patent Publication No. 54-32073, α-glycosyl rutin obtained by this method is prepared by bonding a glucose residue to rutin at equimolar or more in order to improve the water solubility of rutin. It was converted to a rutin glycoside mixture, but did not elucidate the physicochemical properties of each glycoside.

This is clear from the fact that the table on page 3 of the publication states that "the name of the glycoside is presumed, and the position of the new transition of glucose is unknown."

[0013] In exploring the wider use of α-glycosyl rutin, elucidation of the physicochemical properties of each glycoside is expected, and the realization of a novel glycoside having a specified chemical structure is under way. .

[0014]

As described above, the industrialization of a specific novel α-glycosyl rutin whose physicochemical properties have been elucidated is awaited, and its realization has not yet been seen.

[0015]

DISCLOSURE OF THE INVENTION The present invention has been made to solve the above-mentioned problems, and in particular, among α-glycosyl rutin produced by glycosyltransferase,
Focusing on α-glucosyl rutin, which has the smallest molecular weight,
We elucidated its physicochemical properties and studied its production method and application.

As a result, the novel substance 4 represented by the chemical formula 1
^The present invention has been completed by clarifying ^G- α-D-glucopyranosyl rutin, establishing its production method, its use in various compositions, and its antioxidant method.

[0017]

BEST MODE FOR CARRYING OUT THE INVENTION The ^4G- α-D-glucopyranosyl rutin of the present invention has a very high water solubility, which is about 30,000-fold higher than that of rutin, and its molecular extinction coefficient (yellowness ) Is substantially identical to that of rutin,
It has been found that it is an ideal substance having strong antioxidant ability and, moreover, easily hydrolyzed by α-glucosidase in the living body to exert rutin's original physiological effect. In addition, 4 ^{G-.alpha.-D-glucopyranosyl} rutin supersaturated organic solvent solution from the crystallization, by established a process for producing a high-purity 4 ^{G-.alpha.-D-glucopyranosyl} rutin by separating, in a higher purity It also facilitates the production of stable powders that are substantially non-hygroscopic, making their industrial production extremely easy.

The process for producing 4 ^{G-.alpha.-D-glucopyranosyl} rutin of the invention, by the action of a saccharide-transferring enzyme and glucoamylase solution containing rutin and starchy, 4 ^G
It was found that -α-D-glucopyranosyl rutin was produced and collected.

Hereinafter, the present invention will be described in detail.

The rutin used in the present invention is generally desirably purified rutin. If necessary, an extract derived from various plants containing rutin or a partially purified product thereof can be used as appropriate.

Examples of the plant tissue include buckwheat leaf stem,
The buds of A. jujube (Goesseum or Sophoraceae rice), the buds of Echinida, the stalks of eucalyptus, the ginkgo biloba, the citrus fruits, and the like, as well as the tissue cultures of these plants can be advantageously used.

The starch used in the present invention may be any substance capable of producing α-glycosylrutin in which glucose is bound to rutin in equimolar or more by simultaneously used glycosyltransferase, and examples thereof include amylose, dextrin, and the like. Starch partial hydrolysates such as cyclodextrin and maltooligosaccharides, as well as liquefied starch and gelatinized starch are appropriately selected.

Therefore, in order to facilitate the production of α-glycosyl rutin, a starchy substance suitable for the action of glycosyl transfer is selected.

For example, when α-glucosidase (EC 3.2.1.20) is used as a glycosyltransferase, maltooligosaccharides such as maltose, maltotriose, and maltotetraose, or DE of about 10 to 70 are used. A partial hydrolyzate of starch and the like are preferable, and cyclomaltodextrin glucanotransferase (EC 2.4.
When 1.19) is used, cyclodextrin or a partially gelatinized starch having a DE of 1 or less and a partially hydrolyzed starch having a DE of about 60 are preferred. α-amylase (EC3.
When 2.1.1) is used, starch gelatinized products having a DE of 1 or less, dextrin having a DE of about 30 and partially hydrolyzed starch are preferred.

The concentration of starch at the time of the reaction is preferably about 0.5 to 50 times that of rutin.

The solution containing rutin and starch in the present invention preferably contains rutin at a concentration as high as possible.
A solution containing a high concentration in the form of a solution dissolved in an alkaline pH exceeding 7.0 or an aqueous solution of an organic solvent is suitable, and its concentration is about 0.5 W / rutine as rutin.
V% or higher, desirably about 1.0 to 20.0
Solutions containing W / V% are suitable.

[0027] The glycosyltransferase used in the present invention is one which produces α-glycosyl rutin without decomposing rutin when acted on a solution containing rutin and a starchy substance having properties suitable for this enzyme. Good.

For example, α-glucosidase is an enzyme derived from animal or plant tissues such as pig liver and buckwheat seed, or
Mucor, Penicillium
An enzyme derived from a culture obtained by culturing a microorganism such as mold belonging to the genus illium) or yeast belonging to the genus Saccharomyces in a nutrient medium is a cyclomaltodextrin glucanotransferase, a bacterium of the genus Bacillus, Enzymes derived from bacterial cultures belonging to the genus Klebsiella and the like, α-amylase is a bacterium belonging to the genus Bacillus and the like, or Aspergillus (Aspe)
rgillus) can be appropriately selected.

These glycosyltransferases do not necessarily need to be purified and used as long as the above conditions are satisfied.
Usually, the purpose of the present invention can be achieved with a crude enzyme.

If necessary, it may be purified and used by various known methods. Alternatively, a commercially available glycosyltransferase can be used.

The pH and temperature at the time of the reaction may be adjusted so long as the glycosyltransferase acts to generate α-glycosyl rutin.
The pH is selected from 3 to 10, and the temperature is from 10 to 90 ° C.

The amount of enzyme used and the reaction time are closely related, and usually the amount of enzyme is selected so that the reaction is completed in about 5 to 80 hours from the viewpoint of economy.

[0033] The immobilized glycosyltransferase may be used in a batch-wise or continuous manner for the reaction.

Further, if necessary, a tissue of a microorganism, an animal, or a plant having a glycosyltransferase can be cultured in a medium containing starch and rutin to produce α-glycosyl rutin.

The reaction method of the present invention comprises the steps of: adding a solution containing rutin and starch to a glycosyltransferase and glucoamylase (EC);
3.2.1.3) may be applied.

As glucoamylase, those of various origins such as microorganisms and plants can be used. Usually, Aspergillus genus, Rhizopus (Rhi
Commercially available glucoamylase of microbial origin belonging to the genus zopus) can be advantageously used.

In producing ^4G- α-D-glucopyranosyl rutin, a glycosyltransferase and glucoamylase can be allowed to act simultaneously.

Generally, in order to effectively utilize the starch used, α-glycosyl rutin in which a glucose residue has been transferred by equimolar or more by the action of a glycosyltransferase is produced in advance. ^4G
It is desirable to accumulate and produce -α-D-glucopyranosylrutin. In addition, other enzymes such as β-amylase (EC3.
2.1.2), and the combined use of α-L-rhamnosidase and the like are optional.

First, the reaction method of glycosyltransferase will be described.

At the time of the reaction, it is industrially advantageous to carry out the reaction with rutin as high as possible.

For example, when reacting rutin in a high-concentration suspension, about 0.5 W / V% or more, desirably about 1.0 to 5.0 W / V% of suspended rutin and an appropriate amount of rutin are reacted. The rutin-rich solution containing starch is adjusted to a pH of about 4.5 to 6.5, and the temperature is as high as possible at which the glycosyltransferase can act.
Specifically, when the temperature is maintained at about 65 to 90 ° C. and a glycosyltransferase is allowed to act thereon, the suspended rutin gradually dissolves as rutin is converted to α-glycosyl rutin, and at the same time, glucose is added to rutin. Α-Glycosyl rutin bound in equimolar or more is easily formed at a high concentration.

For example, when reacting rutin in the form of a high-concentration solution in which the rutin is dissolved on the alkaline side exceeding pH 7.0, about 0.5 W is added to water having a pH of about 7.5 to 10.0.
/ V% or more, desirably about 1.0 to 5.0 W / V%
Is heated and dissolved, and a rutin-rich solution obtained by dissolving an appropriate amount of starch in the resulting solution is converted to a pH and temperature as high as possible, at which the glycosyltransferase can act, specifically, a pH of about 7.5 to 10 0.0 and a temperature of about 50 to 80 ° C., and when a glycosyltransferase is allowed to act thereon, α-glycosyl rutin is easily produced at a high concentration.

At this time, rutin in the alkaline solution is liable to be decomposed. Therefore, in order to prevent this, it is desirable that rutin be kept under light shielding and anaerobic as much as possible. If necessary, L-
An antioxidant such as ascorbic acid or erythorbic acid may coexist.

For example, when reacting in the form of a high-concentration solution in which rutin is dissolved in an organic solvent aqueous solution, rutin is dissolved in an organic solvent by heating in advance, and then this is mixed with an aqueous starchy solution. , Add glycosyltransferase to this,
Alternatively, when rutin and starch are dissolved in an organic solvent aqueous solution by heating, the mixture is cooled to a predetermined temperature, and a reaction is performed by adding a glycosyltransferase thereto, thereby producing α-glycosyl rutin at a high concentration.

Further, a method for combining the above conditions, for example, a method for preparing a rutin-rich liquid containing about 2.0 to 20.0 W / V% of suspended rutin and an appropriate amount of starchy substance at a pH of about 7.5.
To 10.0 and a temperature of about 50 to 80 ° C., and when a glycosyltransferase is allowed to act thereon, the α-glycosyl
Rutin is easily produced in high concentrations.

As rutin, for example, caustic soda aqueous solution of about 0.1 to 1.0N, caustic potash aqueous solution,
Approximately 5.0 to 20.0 W / in a strongly alkaline aqueous solution such as aqueous sodium carbonate solution, aqueous calcium hydroxide solution, and aqueous ammonia solution.
It is necessary to use a solution dissolved at a high concentration of V%, add an acidic aqueous solution such as hydrochloric acid or sulfuric acid to adjust the pH to a level at which the enzyme can act, add starchy material, and immediately activate the glycosyltransferase. This is extremely advantageous because α-glycosyl rutin can be easily produced at a high concentration.

From the α-glycosyl rutin thus produced by the transglycosylation reaction, the ^4G- α-D of the present invention is obtained.
In order to produce glucopyranosyl rutin, glucoamylase may be allowed to act on the glycosyltransfer reaction solution as it is, or after heat inactivation of the glycosyltransferase. By the action of glucoamylase, the macromolecular substance of α-glycosylrutin undergoes hydrolysis, and D-glucose and ^4G -α-
It accumulates and produces D-glucopyranosyl rutin.

The reaction solution that has produced ^4G- α-D-glucopyranosyl rutin in this manner is usually prepared as ^4G- α-D-glucopyranosyl rutin.
^It contains a relatively small amount of unreacted rutin and a large amount of glucose together with ^G- α-D-glucopyranosyl rutin, and can be directly used as a ^4G- α-D-glucopyranosyl rutin product. Generally, the reaction solution is filtered and concentrated to a syrup-like, and further dried and pulverized to a powdered ^4G- α-D-glucopyranosyl rutin product.

This product is not only a vitamin P enhancer, but also a highly safe natural yellow colorant, antioxidant,
As stabilizers, quality improvers, prophylactics, therapeutics, ultraviolet absorbers, etc., it can be advantageously used in various compositions such as foods and drinks, foods, feeds, feeds, anti-sensitive diseases, cosmetics, and plastic products. . Furthermore, purified ^4G- α-D-
In the case of producing a glucopyranosyl rutin product, for example, the difference in adsorptivity of a porous synthetic adsorbent is used to make a product.
^G- α-D-glucopyranosyl rutin and impurities such as glucose may be separated and purified.

The porous synthetic adsorbent referred to in the present invention is a porous synthetic adsorbent having a large adsorption surface area and a nonionic styrene-divinylbenzene polymer, phenol-formalin resin, acrylate resin, methacrylate resin, and the like. Synthetic resin, for example, commercially available Rohm &
Amberlite XAD-1, Amberlite XAD-2, Amberlite XAD-4 manufactured by Haas
Amberlite XAD-7, Amberlite XAD-
8, Amberlite XAD-11, Amberlite XA
D-12, trade name manufactured by Mitsubishi Kasei Kogyo Co., Ltd. Diaion HP-10, Diaion HP-20, Diaion HP-30, Diaion HP-40, Diaion HP-50, trade name IMACTI manufactured by IMACTI S
yn-42, Imakti Syn-44, Imakti Sy
n-46.

The purification process of the reaction solution yielding 4 ^{G-.alpha.-D-glucopyranosyl} rutin of the invention, the reaction mixture, for example, when passed through a column packed with a synthetic macroporous resin, 4 ^G-.alpha. While -D-glucopyranosyl rutin and a relatively small amount of unreacted rutin are adsorbed to the porous synthetic adsorbent, a large amount of coexisting water-soluble saccharides such as glucose flow out without being adsorbed.

In particular, when the reaction solution contains an organic solvent, the concentration of the organic solvent is reduced, and the solution is brought into contact with a porous synthetic adsorbent. ^G- α-D-glucopyranosyl rutin and rutin may be adsorbed and purified.

If necessary, after the reaction between the glycosyltransferase and the glucoamylase is completed and before contact with the porous synthetic adsorbent, for example, insolubles generated by heating the reaction solution are removed by filtration. Or treated with magnesium aluminate silicate, magnesium aluminate, etc. to adsorb and remove proteinaceous substances in the reaction solution, or use a strongly acidic ion exchange resin (H
Type), basic or weakly basic ion exchange resin (OH
It is optional to use a combination of purification methods such as desalting after treatment with a compound (type).

As described above, a large amount of ^4G- α-D-glucopyranosyl rutin selectively adsorbed on the porous synthetic adsorbent column and a relatively small amount of unreacted rutin are washed with dilute alkali, water or the like. After that, a relatively small amount of an organic solvent or a mixture of an organic solvent and water, for example, methanol water,
First, ^4G- α-D-
Glucopyranosyl rutin is eluted and unreacted rutin is eluted by increasing the flow rate or increasing the concentration of the organic solvent, so this ^4G- α-D-glucopyranosyl rutin-rich fraction, preferably, What is necessary is just to collect a high content fraction having a purity of 75 W / W% or more.

This ^4G- α-D-glucopyranosyl rutin-rich eluate is subjected to a distillation treatment, the organic solvent is first distilled off, and then concentrated to an appropriate concentration to obtain ^4G- α-D-glucopyranosyl rutin. A syrup-like product as a main component is obtained. Further, by drying and pulverizing it, a powdery product containing ^4G- α-D-glucopyranosyl rutin as a main component is obtained.

The elution operation of ^4G- α-D-glucopyranosyl rutin and unreacted rutin with this organic solvent is as follows.
At the same time, the operation of regeneration of the porous synthetic adsorbent is also performed, so that the porous synthetic adsorbent can be used repeatedly.

The purification using the porous synthetic adsorbent of the present invention has a feature that not only water-soluble sugars such as glucose but also impurities such as water-soluble salts can be removed at the same time.

Further, if necessary, in the purification of ^4G- α-D-glucopyranosyl rutin, a method different from the above-mentioned method using a porous synthetic adsorbent, for example, a solubility fractionation method, a molecular weight fractionation method, Employing methods such as membrane separation, high performance liquid chromatography, and column chromatography, or using one or more of these methods in combination with a method using a porous synthetic adsorbent to achieve higher purity The preparation of ^4G- α-D-glucopyranosyl rutin is also optional.

Next, ^4G- α-D-glucopyranosyl
A method for producing crystallization from a supersaturated organic solvent solution of rutin, separating the crystals and producing ^4G- α-D-glucopyranosyl rutin with higher purity will be described.

The ^4G- α-D-glucopyranosyl rutin organic solvent solution for crystallization used in the present invention may be a supersaturated solution, as long as ^4G- α-D-glucopyranosyl rutin is crystallized. ^The method for producing ^G- α-D-glucopyranosyl rutin is not limited.

Usually, the degree of supersaturation is about 1.05 to 1.5, specifically, 4 ^G- α having a purity of 80 W / W% or more.
Preferably, -D-glucopyranosyl rutin is used as an organic solvent solution having a concentration of about 15 to 50 W / W%, and the solution temperature is preferably in a range of 0 to 80 ° C., in which heat loss and organic solvent loss are relatively small in the production process. As the organic solvent solution, for example, methanol, ethanol, acetone or the like is appropriately selected, but it is also optional to use an appropriate amount of water as needed.

In the crystallization method, usually, a supersaturated solution at a relatively high temperature is placed in an auxiliary crystal can, and a seed crystal is desirably added thereto.
A mass kit may be prepared by coexisting the mixture in an amount of 5.0 to 5.0 W / W%, slowly cooling while stirring slowly, and promoting crystallization.

A method for collecting high-purity ^4G- α-D-glucopyranosyl rutin powder from the thus-crystallized mass kit is, for example, by subjecting the mass kit to a basket-type centrifugal separator by a honey separation method, ^4G- α
-D-glucopyranosyl rutin and honey (mother liquor) are separated from each other. If necessary, a small amount of a cold organic solvent or the like can be sprayed on the crystal to wash it, and a higher purity crystalline ^{4G can be obtained.} -Α-D-glucopyranosylrutin was collected, and further dried by heating or the like to completely remove the organic solvent, thereby obtaining high-purity ^4G- α-D-glucopyranosyl.
What is necessary is just to collect rutin powder. The high-purity ^4G- α-D-glucopyranosyl rutin obtained in this manner is substantially non-hygroscopic, fluid, and has a concern of sticking and sticking, although it slightly varies depending on its purity. However, its excellent features are as follows.

(1) The water solubility at room temperature is about 30,000 times higher than that of about 4.3 g dissolved in 1 mL of water and only 1 g of rutin is dissolved in 8 L of water. ing. (2) The molecular extinction coefficient (yellowness) is substantially the same as that of rutin. (3) It has strong antioxidant ability. As an antioxidant, it can be advantageously used to prevent oxidation of oily foods and drinks, oily anti-sensitivity agents, oily cosmetics, and the like. In particular, when used as an anti-sensitivity drug, it exerts its effects as an antioxidant, such as removing active oxygen and suppressing the production of lipid peroxide, and prevents, treats, maintains and promotes various sensitivities. It is convenient. Also, unlike other vitamins and antioxidants such as thiamine, riboflavin, vitamin C and vitamin E,
^4G- α-D-glucopyranosyl rutin is used in combination with one or more vitamins and antioxidants, as it is practically tasteless and odorless and can be used safely without fear of discoloration, browning and off-flavor. It is also convenient. (4) It is hydrolyzed into rutin and glucose by enzymes in the body, and exhibits the original physiological activity (vitamin P) of rutin. The combined use with vitamin C can enhance their biological activity. (5) In the case of a product containing glucose, ^4G -α-
In addition to exerting the effect of D-glucopyranosyl rutin, glucose can exert a shaping, bulking and sweetening effect, and in the case of a purified product from which glucose has been removed, almost shaping and bulking. ^4G- α without
-D-Glucopyranosyl rutin can exert the effect, and is also substantially tasteless and odorless, so it can be seasoned freely.
Can be perfumed.

From these characteristics, ^4G- α-D-glucopyranosyl rutin is a highly safe natural vitamin P
Not only as an enhancer, but alone or in combination with one or more other components, yellow colorants, antioxidants, stabilizers, quality improvers, viral diseases, bacterial diseases, cardiovascular As a preventive agent, therapeutic agent, ultraviolet absorber, etc. for illness, malignant tumors and other susceptible diseases, foods and drinks, foods, feeds, feeds, cosmetics such as anti-sensitive illness agents, skin-skin agents, skin-whitening agents, and various plastic products It can be advantageously used in a composition, preferably in an amount of 0.001 W / W% or more. At this time, together with ^4G- α-D-glucopyranosyl rutin, one or two selected from quercetin, isoquercitrin, rutin, maltosyl rutin, maltotriosyl rutin, maltotetraosyl rutin, maltopentaosyl rutin and the like. Use in combination with the above can also be carried out advantageously.

Further, ^4G- α-D-glucopyranosyl rutin is well harmonized with various substances having tastes such as sourness, salty taste, astringency, umami, and bitterness, and has high acid resistance and heat resistance. General food and drink, favorite foods, for example, soy sauce,
Powdered soy sauce, miso, powdered miso, moromi, hishio, frikake, mayonnaise, dressing, vinegar, three tablespoons vinegar, powdered sushi vinegar, chinese noodles, tentsuyu, noodle soup, sauce, ketchup, grilled meat sauce, curry roux, stew noodles, soup Nonomoto, Dashinomoto, complex seasonings, various flavorings such as mirin, new mirin, table sugar, coffee sugar, rice crackers, hail, rice, kalinto, fertilizer, rice cakes, steamed buns, seaweed, bean jam, sheep, Various Japanese confectionery such as Mizukuri, Nishikitama, jelly, castella, candy, bread, biscuit, cracker, cookie, pie, pudding, cream puff, waffle, sponge cake, donut, chocolate, chewing gum, caramel, candy, etc. , Ice cream, sherbet and other ice confections, fruit syrup pickles, Syrups such as honey, butter cream, custard cream, flour paste, peanut paste, spread, such as fruit paste, pastes, jam, marmalade, syrup pickles,
Fruits such as fructose, processed foods such as vegetables, breads, noodles, cooked rice, cereal processed foods such as artificial meat, salad oils, oils and fats foods such as margarine, pickles such as Fukujin pickles, betta pickles, senmai pickles, and rakkyo pickles Pickles, such as Takuan-zuke-no-zuke and Chinese cabbage-zuke, meat products such as ham and sausage, fish ham, fish sausage, kamaboko, chikuwa, hampen, etc., sea urchin, salted squid, vinegar kelp , Delicacies such as dried sashimi, fugu mirin dried, seaweed foods such as seaweed, wild vegetables, sardines, small fish, shellfish, boiled beans, potato salad, kelp, tempura, etc. Eggs such as dairy drinks, butter, cheese, dairy products, fish meat, livestock meat, fruits, vegetables, etc., bottles, cans, synthetic liquors, brewed sake, fruit liquors, liquors such as Western liquors, coffee, cocoa, jujube Vitamin P enhancer for instant beverages such as tea, black tea, oolong tea, carbonated beverages, lactic acid beverages, lactic acid bacteria beverages, and other soft drinks, pudding mixes, hot cake mixes, instant juices, instant coffee, instant shirako, instant soups, etc. , A yellow colorant, an antioxidant, a quality improving agent, a stabilizer and the like. In addition, vitamin P fortifiers, antioxidants, feeds and feeds for livestock, poultry, bees, silkworms, fish and other raised animals.
It is also convenient to mix and use them for the purpose of improving palatability.

In addition, tobacco, troches, liver oil drops, multivitamin preparations, mouthwashes, mouthwash tablets, mouthwashes, tube feeding agents, oral medicine, injections, toothpaste, lipstick,
Eye shadow, milky lotion, lotion, cream, foundation, sunscreen, shampoo, rinse, etc. various solid, paste, liquid tastes, susceptible disease preventive agent,
The therapeutic agent, ie, an anti-sensitivity disease agent, a skin beautifying agent, can also be advantageously used in combination with cosmetics such as a whitening agent,
Further, it can be advantageously used by blending it with a plastic product or the like as an ultraviolet absorber, a deterioration inhibitor or the like.

Further, the susceptibility disease referred to in the present invention is 4 ^G
-A disease prevented or treated by α-D-glucopyranosyl rutin, such as a viral disease, a bacterial disease, a traumatic disease, an immune disease, rheumatism, diabetes, a cardiovascular disease, a malignant tumor and the like. There may be. The form of the prophylactic or therapeutic agent for ^4G- α-D-glucopyranosyl rutin sensitive disease can be freely selected according to the purpose. For example, sprays, eye drops, nasal drops, gargles, solutions such as injections, ointments, cataplasms, pastes such as creams, powders, granules, capsules,
Solid agents such as tablets. In the preparation, if necessary, combined with one or more other components such as a therapeutic agent, a physiologically active substance, an antibiotic, an auxiliary agent, a bulking agent, a stabilizer, a coloring agent, a flavoring agent and the like. It is also optional.

The dose can be appropriately adjusted depending on the content, administration route, administration frequency and the like. Usually, ^4G -α-
As D-glucopyranosyl rutin, per adult day,
A range from about 0.001 to 50.0 g is preferred.

In the case of cosmetics, it can be used according to the above-mentioned prophylactic and therapeutic agents.

4 As a method of using ^G- α-D-glucopyranosyl rutin, the steps of completing the products of the various compositions include, for example, mixing, kneading, dissolving, dipping, penetrating, spraying, applying, and spraying. A known method such as injection and injection is appropriately selected.

Hereinafter, the ^4G- α-D-glucopyranosyl rutin of the present invention will be described in detail by experiments.

[0073]

[Experiment 1] <4. Preparation of ^G- α-D-glucopyranosyl rutin> 1 part by weight of rutin and dextrin (DE1
2) 10 parts by weight were mixed with 100 parts by weight of hot water at 80 ° C. to obtain a suspension-rich rutin-rich liquid, which was mixed with Bacillus stearot.
20 units per dextrin trams (cyclomaltodextrin glucanotransferase derived from B. thermophilus) were added, and the mixture was reacted at pH 6.0 at 75 ° C. with stirring for 64 hours while stirring. When the reaction solution was analyzed by thin-layer chromatography, most of rutin was converted to α-glycosyl rutin in which glucose residues were bonded in equimolar or more. The reaction solution was heated to inactivate the enzyme, filtered, and the filtrate was concentrated to a concentration of about 30 W / W%, adjusted to pH 5.0, and glucoamylase (EC 3.2.1.3, biochemical 100 units per gram of solid matter)
The reaction was carried out at 5 ° C. for 5 hours. When the reaction solution was analyzed by thin-layer chromatography, a large amount of D-glucose, ^4G- α
-Contains D-glucopyranosyl rutin and a small amount of rutin.

The reaction solution was heated to inactivate the enzyme, filtered, and the filtrate was used as a porous synthetic adsorbent, trade name Diaion HP
-10 (sold by Mitsubishi Chemical Industry Co., Ltd.)
Multiplied by 1.5. The column was passed through with water and washed to remove impurities such as glucose, and then an aqueous ethanol solution was passed through to elute and collect ^4G- α-D-glucopyranosyl rutin and rutin. It has become. This powder is heated and dissolved with hot water to a concentration of about 40 W / W%, a small amount of rutin crystals are added as seed crystals, the mixture is slowly cooled for 2 days with stirring, and centrifuged to remove crystalline rutin. concentrated,
After powdering, ^4G- α-D-glucopyranosyl rutin-rich powder was added to a powder of about 7 wt.
Obtained in 0% yield.

This product contains about 94% of ^4G -α-
It contained D-glucopyranosyl rutin.

[0076]

[Experiment 2] <High purity ^4G- α-D-glucopyranosyl
Preparation of Rutin> The ^4G- α-D-glucopyranosyl rutin powder obtained by the method of Experiment 1 is placed in a glass container, and a concentration of about 35 is added to an aqueous ethanol solution (ethanol: water = 4: 1).
When dissolved by heating to W / W% and kept at room temperature for 5 days, crystals precipitated on the inner wall.

Using these crystals as seed crystals, ^4G- α-D- having a concentration of about 40 W / W% similarly prepared with an aqueous ethanol solution.
The solution was added to the glucopyranosyl rutin solution, and the mixture was subjected to auxiliary crystallization with stirring for 2 days. The resulting mass kit was separated, and the crystals were sprayed with a small amount of an aqueous ethanol solution and washed to obtain crystalline ^4G- α-D-glucopyranosyl rutin. Was.
This product is similarly dissolved in an aqueous ethanol solution, treated, recrystallized, and separated to collect crystalline ^4G- α-D-glucopyranosyl rutin.
9.8 w / w% of ^4G- α-D-glucopyranosyl rutin powder was obtained.

When the physicochemical properties of this powder were examined,
It has been found to be ^4G- α-D-glucopyranosyl rutin, which has never been known before.

Hereinafter, the physicochemical properties of the substance ^4G- α-D-glucopyranosyl rutin of the present invention will be described.

(1) Elemental analysis measured value C = 51.2% H = 5.3% theoretical value C = 51.30% H = 5.22% (molecular formula: C ₃₃ H ₄₀ O ₂₁ ) (2) molecular weight 772.7 (3) Melting point: 232 to 235 ° C (decomposition temperature)

[0081]

(5) Ultraviolet Absorption Spectra The substance of the present invention was dissolved at a concentration of 0.04 mM in acetic acid-ethanol (acetic acid: ethanol = 1: 1,000) and measured. The results are shown in FIG. As a control, rutin was dissolved and measured at the same concentration, and the results are shown in FIG. 1 and 2, the maximum absorption of the substance of the present invention was 362.5 n.
The molecular extinction coefficient at m is ε = 3.50 × 10 ⁴ ,
This value is substantially identical to that of the control rutin. (6) Infrared absorption spectrum Measured by the KBr tablet method. The results are shown in FIG. The infrared absorption spectrum of the substance of the present invention is shown by (I), and that of the control rutin is shown by (II). (7) Solubility in drugs It is easily soluble in water, acetic acid, 0.1N-sodium hydroxide and 0.1N-hydrochloric acid. Soluble in methanol and ethanol. Poorly soluble in chloroform and ethyl acetate. (8) Solubility Dissolve about 4.3 g in 1 mL of water at 25 ° C. In the case of the control rutin, only about 1 g is soluble in 8 L of water. Therefore,
The solubility of the substance of the present invention is more than about 30,000 times that of rutin. (9) Physical properties, color of substance Almost tasteless, odorless yellow powder. Does not deliquesce because it is not hygroscopic. The aqueous solution is neutral to slightly acidic. (10) Stability The stability of the aqueous solution is excellent at pH 3 to 7. (11) Color reaction A green color is obtained by an anthrone-sulfuric acid reaction. Fehring's solution reduction reaction was negative.

(12) Structure (a) Hydrolysis i) Hydrolysis with 1N hydrochloric acid produces 1 mol of L-rhamnose and 2 mol of D-glucose per 1 mol of quercetin. ii) When hydrolyzed with α-glucosidase derived from pig liver, 1 mol of D-glucose is produced for 1 mol of rutin. (B) NMR The substance of the present invention and rutin were measured using an NMR measuring apparatus (VXR-500, manufactured by Varian, USA). The measurement solvent is dimethyl sulfoxide (DMSO)
Was used. The results are summarized in Table 1.

[0084]

[Table 1]

From the results shown in Table 1, a large chemical shift was found only at the carbon at position 4 of the glucose residue of rutin. From the result, the bond of transglycosylated glucose was found to occur in the rutinose residue of the sugar part of rutin. It is judged that the glucose is ether-bonded to the hydroxyl group at the 4-position carbon of the glucose residue.

From the above physicochemical properties, the substance of the present invention is
Novel substance 4 represented by the following chemical formula 1 ^G-Α-D-gluco
It turned out to be pyranosyl rutin.

[0087]

Embedded image

Further, from these results, it can be seen from the results that α-glycosyl in which glucose residues are bonded in equimolar or more to rutin produced by allowing cyclomaltodextrin glucanotransferase to act on a solution containing rutin and starch.
Rutin is determined to have a structure represented by the following chemical formula 3.

[0089]

Embedded image

[0090]

[Experiment 3] <Antioxidant action of ^4G- α-D-glucopyranosyl rutin> As an example of an oily food which is susceptible to oxidation, peanut butter powder is used to give ^4G -α-
The antioxidant action of D-glucopyranosyl rutin was examined.
That is, 83 parts by weight of anhydrous crystalline maltose (manufactured by Hayashibara Co., Ltd., Finetose), 9.6 parts by weight of water and 100 parts by weight of peanut butter were prepared by the method of Reference Example A-4 described later as an antioxidant. High purity ^4G
0.5 parts by weight of α-D-glucopyranosyl rutin powder or 0.5 parts by weight of ^4G- α-D-glucopyranosyl rutin powder and 0.2 parts by weight of citric acid are added, mixed and taken into a vat. Seal and leave at 35 ° C. for 1 day to solidify, pulverize it to obtain peanut butter powder, take it in an aluminum cup, leave it unsealed and leave it in a constant temperature room at 35 ° C. The peroxide value was measured. The peroxide value (meq / Kg) is described in DH Wheeler, Oil and Soap, Volume 9, pages 89 to 97 (1932). It was measured according to the method used. As a control, a group without addition of an antioxidant was provided. The results are shown in Table 2.

[0091]

[Table 2]

As is apparent from the results in Table 2, ^4G- α
It was found that -D-glucopyranosyl rutin has a stronger antioxidant effect than the control, and its antioxidant effect was enhanced by the combined use with citric acid.

[0093]

[Experiment 4] <Acute toxicity> Powdered 4 ^G prepared by the method of Reference Example A-3 described later using a 7-year-old dd mouse.
An acute toxicity test was conducted by oral administration of -α-D-glucopyranosyl rutin product.
No deaths were seen until. Therefore, the toxicity of this substance is extremely low.

Further, when this test was performed using a high-purity ^4G- α-D-glucopyranosyl rutin powder prepared by the method of Reference Example A-4 described later, similar results were obtained.
The toxicity was found to be very low.

Hereinafter, as a reference example of the present invention, ^4G -α-
In Reference Example A, a production example of D-glucopyranosyl rutin was used.
An example of use of ^4G- α-D-glucopyranosyl rutin in various compositions will be described in Reference Example B.

[0096]

Reference Example A-1 < ^4G- α-D-glucopyranosyl
Rutin> 1 part by weight of rutin is heated and suspended in 5 parts by weight of a 50 V / V% ethanol aqueous solution, and separately dextrin (DE8)
Eight parts by weight were dissolved by heating in 45 parts by weight of water, and then these solutions were mixed and mixed with Bacillus stearothermophilus.
ilus) -derived cyclomaltodextrin glucanotransferase (sold by Hayashibara Biochemical Laboratory Co., Ltd.)
Was added to the dextrin per 10 units, adjusted to pH 6.0, and reacted at 50 ° C. for 48 hours. When the reaction solution was analyzed by thin-layer chromatography, it was found that most of the reaction solution was converted to ^4G- α-D-glucopyranosyl rutin having glucose residues bonded in equimolar or more.

The reaction solution was concentrated under reduced pressure to distill off ethanol.
After that, the enzyme was inactivated by heating and adjusted to pH 5.0.
Glucamylase (EC 3.2.1.3, biochemistry)
100 units per gram of solid matter)
Then, the reaction was carried out at 50 ° C. for 5 hours. Further add this reaction solution.
Heat to inactivate the enzyme, filter, and filter the filtrate with ion-exchange resin.
(H type and OH type), desalting, purification and concentration, solid
About 10w / w% of 4 ^G-Α-D-glucopyranosyl
 A small amount of rutin and a large amount of glucose along with rutin
Syrup-like product containing
In a yield of about 95%.

This product is used as a highly water-soluble vitamin P enhancer, and as a highly safe natural yellow colorant, antioxidant, quality improving agent, prophylactic agent, therapeutic agent, ultraviolet absorber, etc. Food and drink, food, feed, feed, anti-sensitivity drugs,
It can be advantageously used for various compositions such as cosmetics and plastic products.

[0099] Since the product contains a large amount of glucose, it can be used as a sweetener.

[0100]

Reference Example A-2 < ^4G- α-D-glucopyranosyl
Rutin> (1) Preparation of α-glucosidase preparation Maltose 4 W / V%, monopotassium phosphate 0.1 W / V
%, Ammonium nitrate 0.1 W / V%, magnesium sulfate 0.05 W / V%, potassium chloride 0.05 W / V%,
Polypeptone 0.2W / V%, calcium carbonate 1W / V
% (Separately by dry heat sterilization and aseptically added at the time of inoculation) and 500 parts by weight of a liquid medium consisting of water to Mucor javanicus IFO4570.
Was shake-cultured at a temperature of 30 ° C. for 44 hours. After completion of the culture, the mycelium was collected, and 0.5 parts with respect to 48 parts by weight of the wet mycelium.
4M urea solution 50 dissolved in M acetate buffer (pH 5.3)
After adding 0 parts by weight, the mixture was allowed to stand at 30 ° C. for 40 hours, and then centrifuged. The supernatant was dialyzed overnight in running water, and then ammonium sulfate was added.
The mixture was left to stand at 4 ° C. overnight at 9 ° C., and the formed salting-out product was collected by filtration, suspended and dissolved in 50 parts by weight of 0.01 M acetate buffer (pH 5.3), and then centrifuged to collect the supernatant. Then, an α-glucosidase preparation was obtained.

(2) ^4G- α-D-glucopyranosyl
Preparation of rutin 4 parts by weight of rutin was dissolved in 40 parts by weight of 0.5N caustic soda solution, adjusted to pH 6.5 with 5N hydrochloric acid solution, and 20 parts by weight of dextrin (DE30) was separately added to 10 parts by weight of water. After heating and dissolving, these solutions were mixed to form a suspended rutin-rich solution, and 10 parts by weight of the α-glucosidase preparation prepared by the method (1) was added thereto.
The reaction was carried out at 55 ° C. for 40 hours while stirring and maintaining at H6.5.

In the same manner as in Reference Example A-1, the reaction solution was heated to deactivate the enzyme, adjusted to pH 5.0, and reacted with glucoamylase. Further, the reaction solution was heated to inactivate the enzyme, filtered, and the filtrate was passed through a column of porous synthetic adsorption resin, Diaion HP-10 (trade name, sold by Mitsubishi Chemical Industry Co., Ltd.) at SV2. . As a result, a large amount of ^4G- α-D-glucopyranosyl rutin and a small amount of unreacted rutin in the solution were adsorbed on the porous synthetic adsorbent, and glucose, salts, etc., flowed out without being adsorbed. Then
The column was passed through and washed with water, and then an aqueous methanol solution was passed through to elute and collect ^4G- α-D-glucopyranosyl rutin and rutin.
A powder product containing 5 w / w% of ^4G- α-D-glucopyranosyl rutin and additionally rutin was obtained in a yield of about 90% based on the weight of rutin as the raw material.

The product is used as a highly water-soluble vitamin P enhancer, and a highly safe natural yellow colorant, antioxidant, stabilizer, quality improver, preventive agent, therapeutic agent, ultraviolet absorber As an anti-deterioration agent, it can be advantageously used for various compositions such as foods and drinks, favorite foods, feeds, feeds, anti-sensitivity agents, cosmetics and plastic products.

[0104]

Reference Example A-3 < ^4G- α-D-glucopyranosyl
Rutin> 1 part by weight of rutin and dextrin (DE1
2) 10 parts by weight were mixed with 98 parts by weight of hot water at 80 ° C. to obtain a suspension-rich rutin-rich solution, and cyclomaltodextrin glucanotransferase was added thereto in an amount of 20 units per dextrin, pH 6.0, and 75 ° C. And maintained for 64 hours with stirring. When the reaction solution was analyzed by thin-layer chromatography, most of the solution was converted to α-glycosylrutin in which glucose residues were bonded in equimolar or more. The reaction solution was heated and inactivated as in Reference Example A-1, adjusted to pH 5.0, and reacted with glucoamylase.

The reaction solution was deactivated by heating, filtered, and the filtrate was passed through a column of a porous synthetic adsorbent, Diaion HP-20 (trade name, sold by Mitsubishi Chemical Industry Co., Ltd.) at SV 1.5. As a result, ^4G- α-D-glucopyranosyl rutin and unreacted rutin in the solution were adsorbed by the porous synthetic adsorbent, and glucose, salts, etc. were flowed out without being adsorbed. Next, the column was passed through with water and washed, and then a methanol aqueous solution was passed through the column to remove ^4G- α-D-glucopyranosyl.
Rutin and rutin were eluted, collected, concentrated under reduced pressure, and powdered. This powder is dissolved by heating with hot water to a concentration of about 50 w / w%, a small amount of rutin crystals are added as seed crystals, and the crystals are subjected to auxiliary crystallization with stirring, centrifuged to remove crystalline rutin, and the supernatant is concentrated. Powdered ^4G powder of about 95 w / w% purity
The -α-D-glucopyranosyl rutin product was obtained in a yield of about 90% based on the weight of the raw rutin per solid.

This product is not only a highly water-soluble vitamin P enhancer, but also a highly safe natural yellow colorant, antioxidant, stabilizer and quality, as in Reference Example A-2. As an improving agent, a preventive agent, a therapeutic agent, an ultraviolet absorber, a deterioration preventing agent, etc., it can be advantageously used for various compositions such as foods and drinks, foods, feeds, feeds, anti-sensitivity agents, cosmetics and plastic products.

[0107]

[Reference Example A-4] <High purity 4^G-Α-D-glucopyrano
Silrutin> 4 obtained by the method of Reference Example A-3^G-Α-D
− Pack the glucopyranosyl rutin product in a glass container
And dissolved by heating with methanol to 25 w / w%.
When kept at room temperature for 3 days, crystals precipitate on the inner wall
Was. The crystals were prepared in the same manner as in 4 ^G-Α-D-gluco
Pyranosyl rutin product concentration about 30w / w% methanol
Solution as seed crystals and slowly stirring for 2 days
Assist the crystallization and dilute the resulting mass kit.
Spray methanol to wash and obtain crystalline 4 ^G
-Α-D-Glucopyranosyl rutin at 70 ° C overnight
Dry in air to obtain a purity of about 99 w / w%^G-Α-D-gluco
Pyranosyl rutin powder was obtained.

This product is a high-purity ^4G- α-D-glucopyranosyl rutin, which is not only a highly water-soluble vitamin P enhancer, but also a highly safe natural yellow colorant and antioxidant. Use as a stabilizer, quality improver, preventive agent, therapeutic agent, ultraviolet absorber, anti-deterioration agent, etc. in various compositions such as foods and drinks, favorite foods, feed, feed, antisensitivity agents, cosmetics, plastic products, etc. It can be used advantageously.

FIG. 4 shows an example of a crystal photograph during crystallization from a methanol solution in this reference example.

Further, this crystal was subjected to a powder X-ray diffractometer (manufactured by Rigaku Denki Co., Ltd., trade name: Gaigerflex RAD-
IIB, using CuKα radiation), the main diffraction angles (2θ) were 4.4 °, 8.4 °, 11.5 °,
18.2 °. In addition, the crystal easily loses these diffraction angles by heating and drying to become an amorphous powder, and thus is considered to be a composite crystal of ^4G- α-D-glucopyranosyl rutin and methanol.

[0111]

Reference Example A-5 < ^4G- α-D-glucopyranosyl
Rutin> 1 part by weight of rutin and dextrin (DE8)
5 parts by weight and 50 parts by weight of water were added, the suspension was heated and the suspended rutin-rich solution dissolved as much as possible was cooled to 75 ° C., and cyclomaltodextrin glucanotransferase was added thereto in an amount of 30 units per dextrin ram, to give a pH of 6.
5. The reaction was carried out for 40 hours while maintaining the temperature at 70 ° C. and stirring.

As in Reference Example A-1, the reaction solution was deactivated with the enzyme, adjusted to pH 5.0, and reacted with glucoamylase. The reaction solution was deactivated by heating, filtered, and the filtrate was purified using a porous synthetic adsorbent, trade name Amberlite XAD-
7 (manufactured by Rohm & Haas)
The solution was passed at 1.5.

As a result, ^4G- α-D-glucopyranosyl rutin and unreacted rutin in the solution were adsorbed on the porous synthetic adsorbent, and glucose, salts, and the like flowed out without being adsorbed.

After the column was passed through and washed with water, the column was passed while increasing the concentration of the aqueous ethanol solution stepwise to give 4 ^G
-A fraction containing a high content of α-D-glucopyranosyl rutin was collected, concentrated and powdered, and a powdery ^4G- α-D-glucopyranosyl rutin product having a purity of about 93 w / w% was added to the weight of the raw material rutin. About 85% yield.

This product is not only a highly water-soluble vitamin P enhancer, but also a highly safe natural yellow colorant, antioxidant, stabilizer and quality, as in Reference Example A-2. It can be advantageously used in various compositions such as foods and drinks, foods, feeds, feeds, anti-sensitivity agents, cosmetics, and plastic products as improvers, preventive agents, therapeutic agents, ultraviolet absorbers, deterioration inhibitors, etc. .

[0116]

Reference Example A-6 <High purity ^4G- α-D-glucopyranosyl rutin> ^4G- α-D obtained by the method of Reference Example A-5
-Glucopyranosyl rutin product was dissolved in an aqueous ethanol solution (ethanol: water = 4: 1) under heating to a concentration of 43 w / w%, and the crystals obtained by the method of Experiment 2 were used as seed crystals to obtain 0.5 w / w. % While stirring slowly.
After auxiliary crystallization for a day, the obtained mass kit is separated, the crystals are washed by spraying a small amount of an aqueous ethanol solution, and the obtained crystalline ^4G- α-D-glucopyranosyl rutin is heated to 80 ° C.
^4G- α-D with a purity of about 98 w / w%
-A glucopyranosyl rutin powder was obtained.

The product is a highly pure ^4G- α-D-glucopyranosyl rutin, which is not only a highly water-soluble vitamin P enhancer, but also a highly safe natural yellow colorant and antioxidant. Use as a stabilizer, quality improver, preventive agent, therapeutic agent, ultraviolet absorber, anti-deterioration agent, etc. in various compositions such as foods and drinks, favorite foods, feed, feed, antisensitivity agents, cosmetics, plastic products, etc. It can be used advantageously.

FIG. 5 shows an example of a crystal photograph during crystallization from an organic solvent solution (ethanol: water = 4: 1) in this reference example.

The crystal was analyzed by a powder X-ray diffractometer in the same manner as in Reference Example A-4.
θ) is 6.8 °, 8.1 °, 15.4 °, 17.8 °
Which is different from the crystals crystallized from methanol obtained in Reference Example A-2.

In addition, these crystals can easily lose these diffraction angles by heating and drying to become amorphous powder.
^It is considered to be a composite crystal of ^G- α-D-glucopyranosyl rutin and ethanol.

[0121]

[Reference Example B-1] <Hard candy> Reduced maltose syrup (marketed by Hayashibara Corporation, registered trademark: Mavit) 1,5
Then, 00 parts by weight were heated and concentrated under reduced pressure until the water content became about 2% or less, to which 15 parts by weight of citric acid and Reference Example A-
3 parts by weight of powdered ^4G- α-D-glucopyranosyl rutin product obtained by the method of 3 and a small amount of lemon flavor,
Then, it was molded and packaged according to a conventional method to obtain a hard candy.

This product is a yellow lemon candy enriched with vitamin P, and has low caries and low calories.

[0123]

[Reference Example B-2] <Boiled Butterfly> Butterfly was peeled, cut into appropriate lengths, immersed in a thin saline solution for several hours, and powdered using the method of Reference Example A-5. The mixture was boiled with a liquid containing a green colorant prepared by blending the α-glycosyl rutin product and Blue No. 1 to obtain boiled green butterflies in water.

[0124] The product not only adds color as a material for various Japanese-style dishes, but also exerts a physiological effect as a dietary fiber.

[0125]

Reference Example B-3 <Fertilization> Water was added to 1 part by weight of waxy seed starch.
2 parts by weight, and while heating gelatinization, add 1.5 g of sugar
Parts by weight, crystalline β-maltose (manufactured by Hayashibara Co., Ltd., San Mart®) 0.7 parts by weight, starch syrup 0.3 parts by weight, and powder ^4G- α-D- obtained by the method of Reference Example A-2 Mix 0.02 parts by weight of glucopyranosyl rutin product,
Thereafter, the fertilizer was manufactured by molding and packaging according to a conventional method.

This product has a good fertilization requirement for both flavor and mouthfeel.
It is a kibi-dango-style Japanese sweet.

[0127]

[Reference Example B-4] <Mixed sweetener> 100 parts by weight of honey, 50 parts by weight of isomerized sugar, 2 parts by weight of brown sugar, and syrup-shaped ^4G- α-D-glucopyranosyl obtained by the method of Reference Example A-1 One part by weight of the rutin product was mixed to obtain a mixed sweetener.

The product is a sweetener enriched with vitamin P and is suitable as a health food.

[0129]

[Reference Example B-5] <Sand cream> Crystalline α-maltose (manufactured by Hayashibara Co., Ltd., registered trademark Finetoose)
1,200 parts by weight, 1,000 parts by weight shortening,
10 parts by weight of a powdery ^4G- α-D-glucopyranosyl rutin product obtained by the method of Reference Example A-5, 1 part by weight of lecithin, 1 part by weight of lemon oil, and 1 part by weight of vanilla oil are mixed by a conventional method and sandwiched. A cream was produced.

This product is a sand cream enriched with vitamin P and colored yellow, prevents oxidation of fats and oils, and has good mouthfeel, melting condition and flavor.

[0131]

[Reference Example B-6] <Orange juice> Orange juice 5
0 parts by weight, 0.1 parts by weight of citric acid, 5 parts by weight of sugar, 0.1 parts by weight of a high-purity ^4G- α-D-glucopyranosyl rutin powder obtained by the method of Reference Example A-6, L-ascorbic acid ( 0.1 part by weight of vitamin C), an appropriate amount of flavor and 46 parts by weight of water were mixed, and then filled and sterilized in a container according to a conventional method to obtain an orange juice product.

This product is an orange juice fortified with vitamin P and vitamin C, and has good color tone and flavor.

[0133]

Reference Example B-7 <Bath agent> DL-sodium lactate 21
Parts by weight, sodium pyruvate 8 parts by weight, Reference Example A-3
^4G- α-D-glucopyranosyl obtained by the method of
5 parts by weight of the rutin product and 40 parts by weight of ethanol were mixed with 26 parts by weight of purified water and an appropriate amount of a coloring agent and a flavor to prepare a bath agent.

The product is suitable as a skin-beautifying agent and a skin-whitening agent, and may be used after being diluted 100 to 10,000 times in hot water for bathing. This product is similar to the case of hot water for bathing,
It can also be advantageously used by diluting it for use in face wash water, lotion and the like.

[0135]

Reference Example B-8 <Emulsion> 0.5 parts by weight of polyoxyethylene behenyl ether, 1 part by weight of polyoxyethylene sorbitol tetraoleate, 1 part by weight of lipophilic glyceryl monostearate, 0.5 part by weight of pyruvic acid 0.5 parts by weight of behenyl alcohol, 1 part by weight of avocado oil, 1 part by weight of high-purity ^4G- α-D-glucopyranosyl rutin powder obtained by the method of Reference Example A-4, and appropriate amounts of vitamin E and a preservative were constantly added. The mixture was dissolved by heating according to the method, and 1 part by weight of sodium L-lactate, 5 parts by weight of 1,3-butylene glycol, 0.1 part by weight of carboxyvinyl ponymer and 85.3 parts by weight of purified water were added, and the mixture was emulsified by a homogenizer. ,
Further, an appropriate amount of a fragrance was added, followed by stirring and mixing to produce an emulsion.

The product has an antioxidant property, is highly stable, and can be advantageously used as a high-quality sunscreen, skin-beautifying agent, skin-whitening agent and the like.

[0137]

Reference Example B-9 <Cream> 2 parts by weight of polyoxyethylene glycol monostearate, 5 parts by weight of self-emulsifying glyceryl monostearate, high purity ^4G- α-D obtained by the method of Reference Example A-6 2 parts by weight of glucopyranosyl rutin powder, 1 part by weight of liquid paraffin, 10 parts by weight of glyceryl trioctanoate and an appropriate amount of a preservative are dissolved by heating according to a conventional method, and 2 parts by weight of L-lactic acid and 1,3-butylene glycol 5 are added thereto. The mixture was added to the mixture by weight, and 66 parts by weight of purified water was added thereto, and the mixture was emulsified with a homogenizer, followed by adding an appropriate amount of a flavor and mixing with stirring to produce a cream.

The product has antioxidant properties, high stability,
It can be advantageously used as a high-quality sunscreen, skin-beautifying agent, and fair-skinning agent.

[0139]

Reference Example B-10 <Antioxidant> 10 parts by weight of a powdery ^4G- α-D-glucopyranosyl rutin product obtained by the method of Reference Example A-3, 2 parts by weight of vitamin E, 0.1 part by weight of lecithin and Mix 0.5 parts by weight of sodium citrate,
An antioxidant was obtained.

This product is formulated in an amount of about 0.01 to 5.0 W / W% in oily foods such as margarine and butter cream; anti-sensitive diseases such as unsaturated fatty acids, oily vitamins and oily hormones; and cosmetics such as emulsions and creams. Thus, it can be advantageously used as an antioxidant, a stabilizer, a quality improving agent and the like.

[0141]

Reference Example B-11 <Antioxidant> 10 parts by weight of the high-purity ^4G- α-D-glucopyranosyl rutin powder obtained by the method of Reference Example A-6 and 0.2 part by weight of sodium citrate were mixed, An antioxidant was obtained.

This product is formulated in an amount of about 0.01 to 5.0 W / W% in oily foods such as margarine and butter cream, anti-sensitive diseases such as unsaturated fatty acids, oily vitamins and oily hormones, and cosmetics such as emulsions and creams. Furthermore, about 0.01 to 2.0 W / W% is blended in foods and drinks containing a natural pigment which is easily discolored, and is advantageously used as an antioxidant, a stabilizer, an anti-fading agent, a quality improving agent and the like. Available.

Hereinafter, embodiments of the present invention will be described.

[0144]

Example 1 <Tablet> 20 parts by weight of ascorbic acid and 13 parts by weight of crystalline β-maltose, 4 parts by weight of corn starch and 3 parts by weight of a powdery α-glucosyl rutin product obtained by the method of Reference Example A-2 were uniformly mixed. 12m in diameter after mixing
Tablets were obtained using an m, 20R punch.

The product is ascorbic acid and ^4G- α-D-
It is a multivitamin with glucopyranosyl rutin. It has good ascorbic acid stability and is easy to swallow.

[0146]

Example 2 <Capsule> Calcium acetate monohydrate 1
0 parts by weight, 50 parts by weight of L-magnesium lactate trihydrate,
57 parts by weight of maltose, 20 parts by weight of a powdery ^4G- α-D-glucopyranosyl rutin product obtained by the method of Reference Example A-3 and 12 parts by weight of a γ-cyclodextrin inclusion compound containing 20% of eicosapentaenoic acid were uniformly mixed. After mixing and granulation by a granulator, the mixture was encapsulated in a gelatin capsule according to a conventional method to produce a capsule containing 150 mg of one capsule.

This product prevents the oxidation of eicosapentaenoic acid, and is used as a high-quality blood cholesterol lowering agent, immunostimulant, skin-beautifying agent, etc. for the prevention and treatment of susceptible diseases,
It can be advantageously used as a food for promoting health.

[0148]

Example 3 <Ointment> 1 part by weight of sodium acetate trihydrate and 4 parts by weight of DL-calcium lactate were uniformly mixed with 10 parts by weight of glycerin, and this mixture was mixed with 50 parts by weight of petrolatum and 10 parts by weight of wax Lanolin, 10 parts by weight, sesame oil 1
4.5 parts by weight, high purity 4 ^G − obtained by the method of Reference Example A-4
An ointment was prepared by adding to a mixture of 1 part by weight of α-D-glucopyranosyl rutin powder and 0.5 part by weight of mentha oil and further uniformly mixing.

The product has antioxidant properties, high stability,
It can be advantageously used as a high-quality sunscreen, beautiful skin agent, fair skin agent, etc., and also as an agent for promoting healing of wounds and burns.

[0150]

Example 4 <Injection> High-purity ^4G- α-D-glucopyranosyl rutin powder obtained by the method of Reference Example A-4 was dissolved in water, purified and filtered according to a conventional method to be pyrogen-free. Transfer the solution to a 20 mL ampoule with ^4G- α-D-
Glucopyranosyl rutin was dispensed to 200 mg, dried under reduced pressure, and sealed to produce an injection.

This injection can be administered alone or as a mixture with other vitamins, minerals, etc., intramuscularly or intravenously. In addition, the product does not need to be stored at low temperature, and has extremely good solubility in physiological saline and the like when used.

This product not only supplements vitamin P, but also acts as an antioxidant, such as removing active oxygen and suppressing the production of lipid peroxide, and is effective for viral diseases, bacterial diseases, cardiovascular diseases, and malignant diseases. It can be advantageously used as a prophylactic or therapeutic agent for various diseases such as tumors.

[0153]

Example 5 <Injection> 6 parts by weight of sodium chloride, 0.3 parts by weight of potassium chloride, 0.2 parts by weight of calcium chloride,
3.1 parts by weight of sodium lactate, 45 parts by weight of maltose, and 2 parts by weight of high-purity ^4G- α-D-glucopyranosyl rutin powder obtained by the method of Reference Example A-6 were dissolved in 1,000 parts by weight of water, Purify and filter according to the method to make it pyrogen-free, and place this solution in a sterilized plastic container.
Injections were prepared by filling each 0 mL.

This product is an injection not only for supplementing vitamin P, but also for supplementing calories and minerals, removing active oxygen, and suppressing the production of lipid peroxide.
Post-illness treatment promotion, recovery promotion, and further viral diseases,
It can be advantageously used as a preventive or therapeutic agent for various diseases such as bacterial diseases, cardiovascular diseases, and malignant tumors.

[0155]

Example 6 <Gavage nutritional supplement> Crystalline α-maltose 20
Parts by weight, 1.1 parts by weight of glycine, 0.18 parts by weight of sodium glutamate, 1.2 parts by weight of sodium chloride, 1 part by weight of citric acid, 0.4 parts by weight of calcium lactate, 0.1 parts by weight of magnesium carbonate, Reference Example A Powder ^4G obtained by the method of -5
0.1 parts by weight of α-D-glucopyranosyl rutin product,
A formulation comprising 0.01 parts by weight of thiamine and 0.01 parts by weight of riboflavin is prepared. Each 24 g of the blend was filled into a laminated aluminum pouch and heat sealed to prepare a tube nutritional supplement.

[0156] The present tube-feeding nutrient can be used for about 300 to 50 bags per bag.
It is dissolved in 0 mL of water, and can be advantageously used as an oral or parenteral nutritional supplement for the nasal cavity, stomach, intestine, etc. by the tube method.

[0157]

As described in the text, the novel substance ^4G- α-D-glucopyranosyl rutin of the present invention can increase the water solubility more than 30,000 times while maintaining the molecular extinction coefficient (yellowness) of rutin. Moreover, it is practically tasteless and odorless, and is easily hydrolyzed in vivo into rutin and D-glucose without any concern about toxicity, and exhibits the original physiological activity of rutin.

In addition, this ^4G- α-D-glucopyranosyl rutin can be easily produced by a biochemical method in which a glycosyltransferase and glucoamylase are allowed to act on a solution containing rutin and starch. It is easy to carry out industrially.

Further, it has been found that the reaction can be carried out by increasing the charged concentration of rutin, and that ^4G- α-D-glucopyranosyl rutin can be easily produced at a high concentration. by also found that the reaction mixture can be purified to 4 ^{G-.alpha.-D-glucopyranosyl} rutin in contact with the porous synthetic adsorbent, 4 ^G -
It facilitates mass production of α-D-glucopyranosyl rutin.

Furthermore, ^4G- α-D-glucopyranosyl
By crystallization and separation from a supersaturated solution of rutin, a method for producing ^4G- α-D-glucopyranosyl rutin having a higher purity has been established, thereby greatly facilitating its industrial realization.

The ^4G -α-
D-glucopyranosyl rutin is not only a highly safe natural vitamin P enhancer, but also a yellow colorant,
Foods, foods, foods, foods, antioxidants, stabilizers, quality improvers, prophylactics, therapeutics, ultraviolet absorbers, anti-deterioration agents, etc.
The composition is advantageously used for feeds, feeds, cosmetics such as anti-sensitivity agents, skin-beautifying agents, fair-skinning agents, and various compositions such as plastic products.

Therefore, the establishment of ^4G- α-D-glucopyranosyl rutin according to the present invention, its industrial production method and its use are of great industrial significance in the food and drink, cosmetics, pharmaceutical and plastic industries.

[Brief description of the drawings]

FIG. 1 is an ultraviolet absorption spectrum of ^4G- α-D-glucopyranosyl rutin of the present invention.

FIG. 2 is an ultraviolet absorption spectrum of a control rutin.

FIG. 3 shows an infrared absorption spectrum, (I) shows an infrared absorption spectrum of the present invention ^4G- α-D-glucopyranosyl rutin, and (II) shows that of a control rutin.

FIG. 4 is a crystal photograph (magnified 40 times) of ^4G- α-D-glucopyranosyl rutin of the present invention crystallized from a methanol solution.

FIG. 5 is a crystal photograph (magnified 40 times) of ^4G- α-D-glucopyranosyl rutin of the present invention crystallized from an ethanol solution.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61K 7/50 A61K 7/50 A61P 3/02 107 A61P 3/02 107 37/00 37/00 // C07H 17/07 C07H 17/07 C12P 19/44 C12P 19/44

Claims (4)

[Claims] [Claim 1] ^4G- α-D- represented by the following chemical formula 1
An anti-sensitive disease agent containing glucopyranosyl rutin. Embedded image 2. The anti-sensitive disease agent according to claim 1, wherein the ^4G- α-D-glucopyranosyl rutin is a crystal. 3. A 4 ^{G-.alpha.-D-glucopyranosyl} rutin, anti-sensitive disease agent according to claim 1 or 2 wherein the purity 75w / w% or more ⁴ G-.alpha.-D-glucopyranosyl rutin. 4. The anti-sensitivity according to any one of claims 1 to 3, further comprising a vitamin selected from thiamine, riboflavin, vitamin C and vitamin E together with ^4G- α-D-glucopyranosyl rutin. Disease agent.