JP3474517B2 - Anti-sensitive disease agent containing 4G-α-D-glucopyranosyl rutin - Google Patents

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an anti-susceptive disease agent containing the novel substance 4 ^G- α-D-glucopyranosyl rutin represented by the following chemical formula 1.

[0002]

[Chemical 2]

[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 vitamin P, which has physiological actions such as capillary strengthening, bleeding prevention, and blood pressure regulation, and as a yellow pigment. It is used for food, medicine, cosmetics, etc.

[0004]

[Chemical 3]

Vitamin P is involved in the biological activity of vitamin C in vivo, for example, the hydroxylation reaction of proline and lysine necessary for the synthesis of collagen which is the main component of living connective tissue, and, for example, cytochrome C. 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 It plays an important role.

[0006] The use of rutin is not limited to vitamin P enhancer as a nutrient, but due to its chemical structure and physiological action, it may be used alone or in combination with other vitamins, for example, a yellow coloring agent, an antioxidant, etc. As food and drink,
In addition, it extends to preventive agents and therapeutic agents for susceptive diseases such as cardiovascular diseases, that is, anti-susceptive agents, and further to cosmetics such as yellowing agents, skin-beautifying agents such as UV absorbers, and skin-whitening agents, and its range is extremely wide. .

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

As a method for improving this, for example, as shown in Japanese Patent Publication No. 25-1677, a method for increasing 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 Publication (Kokai) No. 1989-163, a method of reacting rutin with a monohalogen acetic acid to form a sodium acetate compound to increase water solubility, and Japanese Patent Publication No.
As disclosed in Japanese Patent No. 1285, there has been known a method in which rugalit is allowed to act on rutin to form a sulfite compound to increase water solubility.

However, in all of these methods, an amino compound, a monohaloacetic acid, a sulfite compound, etc. are used, and there is a concern that the product may have other physiological activities and toxicity.
Further, its purification is also difficult.

Therefore, the inventors of the present invention have previously proposed Japanese Patent Publication No.
In JP-32073-A, as a method of water solubilization utilizing a safer biosynthetic reaction, an equimolar amount or more of a glucose residue is transferred from a starch partial hydrolyzate to rutin by the action of a glycosyltransferase, and α- We proposed a method for producing glycosyl rutin and improving its water solubility.

The α-glycosyl rutin obtained by this method, as described in JP-B-54-32073, has a glucose residue bound to rutin in an equimolar amount or more in order to improve the water solubility of rutin. It has been converted to a rutin glycoside mixture, and the physicochemical properties of each glycoside have not been elucidated.

This is also apparent from the fact that the table on page 3 of the same publication states that "the glycoside name is a presumption and the position of new transition of glucose is unknown."

In order to develop a wider range of applications of α-glycosyl rutin, the physicochemical properties of each glycoside are awaited to be elucidated, and the realization of a new glycoside having a specified chemical structure has been sought. .

[0014]

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

[0015]

The present invention has been made to solve the above-mentioned problems, and in particular, among α-glycosyl rutins produced by glycosyltransferase,
Focusing on α-glucosyl rutin, which has the smallest molecular weight,
The physicochemical properties were elucidated, and the manufacturing method and the application were intensively studied.

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 and establishing its production method, various compositions, and its use in an antioxidant method.

[0017]

BEST MODE FOR CARRYING OUT THE INVENTION The 4 ^G- α-D-glucopyranosyl rutin of the present invention has extremely high water solubility, which is about 30,000 times 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 that has a strong antioxidant ability and, moreover, is easily hydrolyzed by α-glucosidase in the living body to exert the original physiological effect of rutin. 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 a substantially non-hygroscopic and stable powder, which greatly facilitates its industrial production.

The method for producing 4 ^G- α-D-glucopyranosyl rutin of the present invention comprises reacting a solution containing rutin and starch with a glycosyltransferase and glucoamylase to produce 4 ^G
It was found that it was sufficient to produce -α-D-glucopyranosyl rutin and collect this.

The present invention will be described in detail below.

The rutin used in the present invention is generally purified rutin, and if necessary, various plant-derived extracts containing rutin, or partially purified products thereof can be used as appropriate.

Examples of plant tissues include buckwheat leaf stems,
The buds of Enju (camellia japonica or succulent rice), the buds of gall, eucalyptus leaves, ginkgo leaves, citrus fruits, and the like, and tissue cultures of these plants can be advantageously used.

The starch used in the present invention may be any one that can produce α-glycosyl rutin in which glucose residues are bound to rutin in an equimolar amount or more by a glycosyltransferase used at the same time, and examples thereof include amylose and dextrin. A partial hydrolyzate of starch such as cyclodextrin and maltooligosaccharide, and further, liquefied starch, gelatinized starch and the like are appropriately selected.

Therefore, in order to facilitate the production of α-glycosyl rutin, a starch material suitable for the action of transglycosylation is selected.

For example, when α-glucosidase (EC 3.2.1.20) is used as a glycosyltransferase, a maltooligosaccharide such as maltose, maltotriose, maltotetraose, or DE of about 10 to 70 is used. Partial starch hydrolysates are preferable, and cyclomaltodextrin glucanotransferase (EC 2.4.
When 1.19) is used, cyclodextrin or a starch gelatinization product having a DE of 1 or less to a starch partial hydrolyzate having a DE of about 60 is suitable. α-amylase (EC3.
When 2.1.1) is used, a starch gelatinized product having a DE of 1 or less, a dextrin having a DE of about 30 or a starch partial hydrolyzate is suitable.

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

The solution containing rutin and starch in the present invention is preferably a solution containing rutin in a concentration as high as possible, for example, in a suspension form at a high concentration or at pH.
A solution containing a high concentration in the form of a solution dissolved at an alkaline pH exceeding 7.0 or dissolved in an organic solvent aqueous solution is suitable, and the concentration thereof is about 0.5 W / rutin.
High concentration of V% or more, preferably about 1.0 to 20.0
A solution containing W / V% is suitable.

The glycosyltransferase used in the present invention can be any one as long as it produces α-glycosyl rutin without degrading rutin when it is reacted with a solution containing rutin and a starchy substance having suitable properties for this enzyme. Good.

[0028] For example, α-glucosidase is an enzyme derived from animal or plant tissues such as pig liver and buckwheat seed, or
Mucor genus, Penicillium (Penic)
The enzyme derived from the culture obtained by culturing a microorganism such as a fungus belonging to the genus Ilium) or a yeast belonging to the genus Saccharomyces in a nutrient medium is cyclomaltodextrin glucanotransferase, and is a bacterium of the genus Bacillus (Bacillus), The enzyme derived from a bacterial culture belonging to the genus Klebsiella or the like is α-amylase, which is a bacterium belonging to the genus Bacillus or the like, or Aspergillus (Aspe).
Enzymes derived from fungal cultures belonging to the genus r.

These glycosyltransferases do not necessarily need to be used after being purified as long as the above conditions are satisfied.
Usually, a crude enzyme can achieve the object of the present invention.

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

The pH and temperature during the reaction may be such that a glycosyltransferase acts to produce α-glycosyl rutin, and usually,
It is selected from pH 3 to 10 and temperature 10 to 90 ° C.

There is a close relationship between the amount of enzyme used and the reaction time, and usually the amount of enzyme is selected so that the reaction is completed in about 5 to 80 hours from the economical point of view.

It is also possible to appropriately select whether the immobilized glycosyltransferase is used in the reaction in a batch system or in a continuous system.

Further, if necessary, the tissue of a microorganism, animal or 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 glycosyltransferase and glucoamylase (EC) to a solution containing rutin and starch.
3.2.1.3) may be applied.

As the glucoamylase, those of various origins such as microorganisms and plants can be used. Usually, Aspergillus genus, Rhizopus (Rhi
Commercially available glucoamylase derived from a microorganism belonging to the genus Zopus) can be advantageously used.

When producing 4 ^G- α-D-glucopyranosyl rutin, a glycosyltransferase and glucoamylase can be simultaneously acted on.

Generally, in order to effectively utilize the starch material to be used, a glycosyltransferase is allowed to act in advance to produce α-glycosyl rutin in which glucose residues have been transferred in an equimolar amount or more, and then glucoamylase. 4 ^G
It is desirable to accumulate and produce -α-D-glucopyranosyl rutin. In addition to glucoamylase, other enzymes such as β-amylase (EC 3.
It is optional to use 2.1.2), α-L-rhamnosidase and the like together.

First, the reaction method of glycosyltransferase will be described.

In the reaction, it is industrially advantageous to incorporate rutin in a concentration as high as possible.

For example, when rutin is reacted in a high-concentration suspension, the amount of suspended rutin of about 0.5 W / V% or more, preferably about 1.0 to 5.0 W / V%, and an appropriate amount. The pH of the rutin-rich liquid containing starch is about 4.5 to 6.5, and the temperature is as high as possible for the action of the glycosyltransferase.
Specifically, when the temperature is maintained at about 65 to 90 ° C. and a glycosyltransferase is allowed to act on this, 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 an equimolar amount or more is easily produced in high concentration.

When rutin is reacted in the form of a high-concentration solution prepared by dissolving rutin 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, preferably about 1.0 to 5.0 W / V%
Solution containing a high amount of rutin, which is obtained by dissolving and dissolving a suitable amount of starchy substance, is prepared at a pH as high as possible for the action of a glycosyltransferase, at a high temperature, specifically, at a pH of about 7.5 to 10. When the temperature is maintained at 0.0 and the temperature is about 50 to 80 ° C. and a glycosyltransferase acts on this, α-glycosyl rutin is easily produced in a high concentration.

At this time, since rutin in the alkaline solution is prone to be decomposed, it is desirable to keep it shielded from light and kept under anaerobic condition as much as possible in order to prevent the decomposition. If necessary, L-
An antioxidant such as ascorbic acid or erythorbic acid may coexist.

For example, in the case of reacting rutin in a high-concentration solution prepared by dissolving it in an organic solvent aqueous solution, rutin is heated and dissolved in an organic solvent in advance, and then this is mixed with an aqueous starchy solution. , Add glycosyltransferase to it,
Alternatively, rutin and a starchy substance are heated and dissolved in an aqueous solution of an organic solvent, cooled to a predetermined temperature, and reacted by adding a glycosyltransferase to the α-glycosyl rutin at a high concentration.

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

As rutin, for example, caustic soda aqueous solution, caustic potash aqueous solution of about 0.1 to 1.0 N,
Approximately 5.0 to 20.0 W / in strong alkaline aqueous solution such as sodium carbonate aqueous solution, calcium hydroxide aqueous solution, and ammonia water.
Using a solution dissolved in a high concentration of V% and adding an acidic aqueous solution of hydrochloric acid, sulfuric acid or the like to this to adjust to a pH at which the enzyme can act, and adding starch and immediately acting a glycosyltransferase, This is very convenient because it easily produces α-glycosyl rutin in high concentration.

From the α-glycosyl rutin thus produced by the transglycosylation reaction, the 4 ^G- α-D of the present invention can be obtained.
-To produce glucopyranosyl rutin, glucoamylase may be allowed to act on the transglycosylation reaction solution as it is, or after deactivating the glycosyltransferase by heating. Due to the action of glucoamylase, the high-molecular substance of α-glycosyl rutin is hydrolyzed, and D-glucose and 4 ^G -α-
Accumulates and produces D-glucopyranosyl rutin.

The reaction solution thus produced with 4 ^G -α-D-glucopyranosyl rutin is usually 4
^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 4 ^G- α-D-glucopyranosyl rutin product. Generally, the reaction solution is filtered and concentrated to give a syrup-like product, and further dried and pulverized into a powdery 4 ^G- α-D-glucopyranosyl rutin product.

This product is not only a vitamin P enhancer but also a highly safe natural type yellow colorant, antioxidant,
As stabilizer, quality improver, preventive agent, therapeutic agent, ultraviolet absorber, etc., it can be advantageously used for various compositions such as food and drink, favorite food, feed, feed, anti-susceptive disease agent, cosmetics and plastic products. . Furthermore, purified 4 ^G- α-D-
When manufacturing a glucopyranosyl rutin product, for example, it is necessary to
^G- α-D-glucopyranosyl rutin and impurities such as glucose may be separated and purified.

The term "porous synthetic adsorbent" as used in the present invention means a nonionic styrene-divinylbenzene polymer, phenol-formalin resin, acrylate resin, methacrylate resin or the like which is porous and has a large adsorption surface area. Synthetic resin such as commercially available Rohm &
Amberlite XAD-1, Amberlite XAD-2, Amberlite XAD-4, trade names manufactured by Haas
Amberlite XAD-7, Amberlite XAD-
8, Amberlite XAD-11, Amberlite XA
D-12, product name manufactured by Mitsubishi Kasei Kogyo Co., Ltd. Diaion HP-10, Diaion HP-20, Diaion HP-30, Diaion HP-40, Diaion HP-50, product name Imacti manufactured by IMACTI S
yn-42, Imacty Syn-44, Imacty Sy
n-46 and the like.

The method for purifying the reaction solution in which 4 ^G- α-D-glucopyranosyl rutin of the present invention is produced is such that when the reaction solution is passed through a column packed with a porous synthetic adsorbent, for example, 4 ^G- α While -D-glucopyranosyl rutin and a relatively small amount of unreacted rutin are adsorbed on 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 and the porous synthetic adsorbent are brought into contact with each other. ^G- α-D-glucopyranosyl rutin and rutin may be adsorbed and purified.

If necessary, after the reaction between the glycosyltransferase and glucoamylase is completed and before contact with the porous synthetic adsorbent, for example, insoluble matter generated by heating the reaction solution is filtered off. Or, it may be treated with magnesium aluminate silicate, magnesium aluminate, etc. to adsorb and remove the proteinaceous substances in the reaction solution, and the strong acid ion exchange resin (H
Type), medium basic or weak basic ion exchange resin (OH
It is optional to use a combination of purification methods such as desalting by treating with a type).

As described above, a large amount of 4 ^G- α-D-glucopyranosyl rutin selectively adsorbed on the porous synthetic adsorbent column and a relatively small amount of unreacted rutin were washed with a dilute alkali, water or the like. After that, a relatively small amount of an organic solvent or a mixed solution of an organic solvent and water, for example, methanol water,
If ethanol water or the like is passed through, 4 ^G- α-D-
Glucopyranosylrutin is eluted, and unreacted rutin is eluted by increasing the flow rate or increasing the organic solvent concentration. Therefore, this 4 ^G- α-D-glucopyranosyl rutin-rich fraction, desirably, A high content fraction having a purity of 75 W / W% or more may be collected.

This ^4G- α-D-glucopyranosylrutin-rich eluate is subjected to a distillation treatment to distill off the organic solvent first and then to a suitable concentration to give ^4G- α-D-glucopyranosylrutin. A syrup-like product containing the main component is obtained. Further, by drying and pulverizing this, a powdered product containing 4 ^G- α-D-glucopyranosyl rutin as a main component is obtained.

The elution operation of 4 ^G -α-D-glucopyranosyl rutin and unreacted rutin with this organic solvent is as follows.
At the same time, it also serves to regenerate the porous synthetic adsorbent, which enables repeated use of the porous synthetic adsorbent.

Further, the purification by 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.

If necessary, in purifying 4 ^G- α-D-glucopyranosyl rutin, a method different from the method using the above-mentioned porous synthetic adsorbent, for example, the solubility fractionation method, the molecular weight fractionation method, A method such as a membrane separation method, high performance liquid chromatography, or column chromatography is adopted, or one or more of these methods are used in combination with a method using a porous synthetic adsorbent to further improve the purity. Of 4 ^G- α-D-glucopyranosyl rutin is also optional.

Next, 4 ^G -α-D-glucopyranosyl
A method of crystallizing rutin from a supersaturated organic solvent solution and separating the crystals to produce further highly pure ^4G- α-D-glucopyranosyl rutin will be described.

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

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

In the crystallization method, usually, a relatively high temperature supersaturated liquid is placed in a supporting crystal can, and seed crystals are preferably added to the auxiliary crystal can at 0.1.
To 5.0 W / W% and co-exist, and slowly cooled with slow stirring to promote crystallization and form a mass kit.

The method of collecting the high-purity 4 ^G- α-D-glucopyranosyl rutin powder from the crystallized musket in this way is, for example, a honey method, and the musket is put into a basket type centrifuge to crystallize. 4 ^G- α
In the method of separating the -D- glucopyranosyl rutin and honey (mother liquor), if necessary, it is easy to wash by spraying a small amount of cold organic solvent to the crystalline products, higher-purity crystalline 4 ^G -Α-D-glucopyranosyl rutin was collected and further dried by heating to completely remove the organic solvent, thereby obtaining 4 ^G- α-D-glucopyranosyl of high purity.
Collect rutin powder. The high-purity 4 ^G- α-D-glucopyranosyl rutin obtained in this manner is substantially non-hygroscopic, fluid, and may cause sticking or sticking, although it varies somewhat depending on its purity. However, its excellent features are as follows.

(1) The water solubility at room temperature was improved by about 30,000 times or more, compared with the fact that about 4.3 g of water was dissolved in 1 mL of water and only 1 g of rutin was dissolved in 8 L of water. ing. (2) The molecular extinction coefficient (yellowness) is substantially the same as that of rutin. (3) It has a strong antioxidant ability. As an antioxidant, it can be advantageously used for preventing oxidation of oily foods and drinks, oily anti-susceptive diseases, oily cosmetics and the like. In particular, when used as an anti-susceptive disease agent, it exerts effects such as removal of active oxygen and suppression of lipid peroxide production as an antioxidant, and prevents, treats, maintains health and promotes various susceptible diseases. It is convenient for Also, unlike other vitamins and antioxidants such as thiamine, riboflavin, vitamin C, vitamin E,
It is virtually tasteless and odorless, and can be used with peace of mind without fear of discoloration, browning or offensive odor. Use 4 ^G- α-D-glucopyranosyl rutin in combination with one or more vitamins and / or antioxidants. It is also convenient. (4) It is hydrolyzed into rutin and glucose by an enzyme in the body, and exhibits rutin's original physiological activity (vitamin P). When used in combination with vitamin C, their physiological activity can be enhanced. (5) In the case of a product containing glucose, 4 ^G -α-
Not only the effect of D-glucopyranosyl rutin is exerted, but also glucose is capable of exerting a shaping effect, an increasing effect and a sweetening effect, and in the case of a purified product from which glucose is removed, almost shaping and increasing the effect. Without 4 ^G- α
-The effect of D-glucopyranosyl rutin can be exhibited, and since it is virtually tasteless and odorless, it can be seasoned freely.
You can perfume.

From these characteristics, 4 ^G- α-D-glucopyranosyl rutin is a highly safe natural vitamin P.
Not only as a strengthening agent, but also alone or in combination with one or more other components, a yellow coloring agent, an antioxidant, a stabilizer, a quality improving agent, a viral disease, a bacterial disease, a circulatory organ. As a preventive agent, therapeutic agent, ultraviolet absorber, etc. for susceptive diseases such as diseases and malignant tumors, foods and drinks, favorite foods, feeds, foods, anti-susceptive disease agents, skin-care agents such as skin-whitening agents, and plastic products. It can be advantageously used by blending it in the composition, preferably 0.001 W / W% or more. In this case, one or two kinds selected from quercetin, isoquercitrin, rutin, maltosylrutin, maltotriosylrutin, maltotetraosylrutin, maltopentaosylrutin together with ^4G- α-D-glucopyranosylrutin. It can also be advantageously used in combination with the above.

Further, 4 ^G- α-D-glucopyranosyl rutin is in good harmony with various substances having tastes such as sourness, salt to taste, astringency, umami, bitterness, etc., and is also highly resistant to acid and heat. General food and drink, favorite foods such as soy sauce,
Powdered soy sauce, miso, powdered miso, moromi, hisoio, furikake, mayonnaise, dressing, vinegar, sancha vinegar, powdered sushi vinegar, Chinese cabbage, tempura sauce, noodle sauce, sauce, ketchup, grilled meat sauce, curry roux, stew sauce, soup Nomoto, Dashi, compound seasoning, various seasonings such as mirin, new mirin, table sugar, coffee sugar, rice crackers, hail, okono, carinto, fertilizer, rice cakes, steamed bun, uiro, bean paste, yogurt, Various Japanese sweets such as Mizuyomi, Nishikidama, jelly, castella, candy, etc., Western sweets such as bread, biscuits, crackers, cookies, pies, pudding, cream puffs, waffles, sponge cakes, donuts, chocolate, chewing gum, caramel, candy. , Ice cream, sherbet and other frozen desserts, 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 of vegetables, breads, noodles, cooked rice, processed foods of cereals such as artificial meat, salad oil, fats and oils foods such as margarine, pickles such as Fukujin pickles, betara pickles, Senmai pickles and rakkyo pickles. , Pickled vegetables such as Takuan pickles and Chinese cabbage pickles, Meat products such as ham and sausages, Fish ham, Fish sausages, Fish products such as kamaboko, chikuwa and hampen, sea urchin and squid salt, vinegar kelp , Various kinds of delicacies such as Saki simmer, dried pufferfish, dried seaweed, edible wild plants, sardines, small fish, seafood such as boiled beans, potato salad, kelp rolls, tempura, Kinshi egg, Milk drinks, butter, eggs such as cheese, dairy products, bottled products such as fish meat, meat, fruits and vegetables, canned products, synthetic liquor, sake, alcohol such as fruit liquor, Western liquor, coffee, cocoa, jue Vitamin P enhancer for instant drinks such as tea, tea, oolong tea, carbonated drinks, lactic acid drinks, lactic acid bacteria drinks, pudding mix, hot cake mix, instant juice, instant coffee, instant shiruko, instant soup, etc. , Yellow colorants, antioxidants, quality improvers, stabilizers and the like. In addition, for feeds and feeds for domestic animals such as livestock, poultry, bees, silkworms, and fish, vitamin P enhancers, antioxidants,
It is also convenient to mix and use it for the purpose of improving palatability.

In addition, tobacco, troches, liver oil drops, complex vitamin preparations, mouth fresheners, oral tablets, mouthwash, tube feeding, oral medication, injections, toothpaste, lipstick,
Eyeshadow, milky lotion, lotion, cream, foundation, sunscreen, shampoo, conditioner, etc.
It can also be advantageously used by being mixed with a therapeutic agent, that is, an anti-susceptive disease agent, a skin-beautifying agent, a cosmetic such as a skin-whitening agent, and the like.
Further, it can be advantageously used by blending it with a plastic product or the like as an ultraviolet absorber, a deterioration preventing agent or the like.

The susceptive diseases referred to in the present invention are 4 ^G
-A disease that is prevented or treated by α-D-glucopyranosyl rutin, such as viral diseases, bacterial diseases, traumatic diseases, immune diseases, rheumatism, diabetes, cardiovascular diseases, malignant tumors, etc. It may be. The shape of the preventive and therapeutic agents for 4 ^G- α-D-glucopyranosyl rutin-sensitive diseases can be freely selected according to the purpose. For example, sprays, eye drops, nasal drops, mouthwash, liquids such as injections, ointments, plasters, pastes such as creams, powders, granules, capsules,
It is a solid preparation such as tablets. In the preparation, if necessary, in combination with other components, for example, one or more of therapeutic agents, physiologically active substances, antibiotics, adjuvants, extenders, stabilizers, coloring agents, flavoring agents and the like. It is also optional to do so.

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

In the case of cosmetics as well, it can be used according to the preventive agent and therapeutic agent described above.

The method of using 4 ^G- α-D-glucopyranosyl rutin includes, for example, mixing, kneading, dissolving, dipping, penetrating, spraying, coating and spraying in the steps until the products of various compositions are completed. A known method such as injection can be appropriately selected.

The 4 ^G -α-D-glucopyranosyl rutin of the present invention will be described in detail below in experiments.

[0073]

[Experiment 1] <Preparation of 4 ^G- α-D-glucopyranosyl rutin> 1 part by weight of rutin and dextrin (DE1)
2) 10 parts by weight was mixed with 100 parts by weight of hot water at 80 ° C. to prepare a suspension-like liquid having a high content of rutin, which was mixed with Bacillus stearothermophilus.
Cyclomaltodextrin glucanotransferase derived from Hermophilus (sold by Hayashibara Biochemical Laboratories, Inc.) was added in an amount of 20 units per dextrin grum, and the mixture was allowed to react for 64 hours while stirring at pH 6.0 and 75 ° C. When the reaction solution was analyzed by thin layer chromatography, most of rutin was converted to α-glycosyl rutin having glucose residues bonded in equimolar amounts or more. The reaction solution is heated to deactivate the enzyme, filtered, and the filtrate is concentrated to a concentration of about 30 W / W% and adjusted to pH 5.0, to which glucoamylase (EC 3.2.1.3, biochemistry). Sold by Kogyo Co., Ltd.) and added 100 units per gram of solid matter,
The reaction was carried out at ℃ for 5 hours. When the reaction solution was analyzed by thin layer chromatography, a large amount of D-glucose, 4 ^G- α
It contained -D-glucopyranosyl rutin and a small amount of rutin.

The reaction solution is heated to inactivate the enzyme, filtered, and the filtrate is a porous synthetic adsorbent, trade name Diaion HP.
SV on the column of -10 (sold by Mitsubishi Kasei Co., Ltd.)
I played with 1.5. After passing through this column with water and washing to remove impurities such as glucose, an aqueous ethanol solution was passed through to elute and collect 4 ^G- α-D-glucopyranosyl rutin and rutin, concentrate under reduced pressure, and powder. Turned into This powder was heated and dissolved in hot water to a concentration of about 40 W / W%, and a small amount of rutin crystals was added as seed crystals, and the mixture was gradually cooled with stirring for 2 days and centrifuged to remove the crystalline rutin, and the supernatant was removed. concentrated,
Pulverize the powder containing 4 ^G- α-D-glucopyranosyl rutin to a high content of about 7 relative to the weight of rutin of the raw material per solid matter.
Obtained in a yield of 0%.

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

[0076]

[Experiment 2] <High-purity 4 ^G- α-D-glucopyranosyl
Preparation of rutin> 4 ^G- α-D-glucopyranosyl rutin powder obtained by the method of Experiment 1 was placed in a glass container, and the concentration thereof was about 35 with an aqueous ethanol solution (ethanol: water = 4: 1).
It was dissolved by heating so as to be W / W% and kept at room temperature for 5 days, and crystals were deposited on its inner wall.

Using this crystal as a seed crystal, 4 ^G- α-D-was similarly prepared with an aqueous ethanol solution and having a concentration of about 40 W / W%.
Add to the glucopyranosyl rutin solution, support the crystals for 2 days with slow stirring, separate the resulting mass kit, spray the crystals with a small amount of an aqueous ethanol solution and wash to obtain crystalline ^4G- α-D-glucopyranosyl rutin. It was
Similarly, this product is dissolved in an aqueous ethanol solution, treated, recrystallized, and concentrated to collect crystalline ^4G- α-D-glucopyranosylrutin, which is vacuum dried overnight at 80 ° C to obtain a purity of 9
9.8 w / w% of 4 ^G- α-D-glucopyranosyl rutin powder was obtained.

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

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

(1) Elemental analysis Measurement 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 spectrum The substance of the present invention was dissolved in acidic acetic acid ethanol (acetic acid: ethanol = 1: 1,000) at a concentration of 0.04 mM for measurement. The results are shown in Figure 1. As a control, rutin was dissolved and measured at the same concentration, and the results are shown in FIG. From the results of FIGS. 1 and 2, the maximum absorption of the substance of the present invention was 362.5n.
The molecular extinction coefficient at m is ε = 3.50 × 10 ⁴ ,
This value is substantially the same as that of the control rutin. (6) Infrared absorption spectrum It was measured by the KBr tablet method. The results are shown in Figure 3. 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 medicine Easily soluble in water, acetic acid, 0.1N-sodium hydroxide, 0.1N-hydrochloric acid. Soluble in methanol and ethanol. Insoluble in chloroform and ethyl acetate. (8) Solubility Approximately 4.3 g is dissolved 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 substance of the present invention has the solubility improved about 30,000 times or more as compared with the case of rutin. (9) Physical properties, substance color, almost no taste and odorless yellow powder. It is not hygroscopic and does not deliquesce. 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 produced by anthrone-sulfuric acid reaction. Fehling's solution reduction reaction is negative.

(12) Structure (a) Hydrolysis i) When hydrolyzed with 1N-hydrochloric acid, 1 mol of L-rhamnose and 2 mol of D-glucose are produced with respect to 1 mol of quercetin. ii) When hydrolyzed with α-glucosidase derived from porcine liver, 1 mol of D-glucose is produced with respect to 1 mol of rutin. (B) NMR The substance of the present invention and rutin were measured using an NMR measuring device (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 in Table 1, it was found that a large chemical shift was found only in the 4-position carbon of the glucose residue of rutin. It is judged that an ether bond is formed 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 was found to be pyranosyl rutin.

[0087]

[Chemical 4]

Further, from these results, α-glycosyl having glucose residues in equimolar amounts or more bound to rutin produced by the action of cyclomaltodextrin glucanotransferase on a solution containing rutin and starch.
Rutin is considered to have a structure represented by Chemical Formula 3 below.

[0089]

[Chemical 5]

[0090]

[Experiment 3] <4 ^G- α-D-glucopyranosyl rutin's antioxidative action> As an example of an oily food susceptible to oxidation, peanut butter powder was used to give 4 ^G -α-
The antioxidant effect of D-glucopyranosyl rutin was examined.
That is, 100 parts by weight of peanut butter, 83 parts by weight of anhydrous crystalline maltose (produced by Hayashibara Co., Ltd., registered trademark FINETOSE), 9.6 parts by weight of water, and an antioxidant were prepared by the method of Reference Example A-4 described later. High purity 4 ^G
0.5 part by weight of -α-D-glucopyranosyl rutin powder or its 4 ^G- α-D-glucopyranosyl rutin powder 0.5 part by weight and 0.2 part by weight of citric acid were added and mixed, and collected in a vat. Seal and leave at 35 ° C for 1 day to solidify, powderize this to obtain peanut butter powder, take this in an aluminum cup, leave it unsealed in a 35 ° C constant temperature room, The peroxide value was measured. The peroxide value (meq / Kg) is described in DH Wheeler, Oil and Soap, Volume 9, pp. 89-97 (1932). It measured according to the method. As a control, an antioxidant-free section was provided. The results are shown in Table 2.

[0091]

[Table 2]

As is clear from the results of Table 2, 4 ^G- α
It was found that -D-glucopyranosyl rutin has a stronger antioxidant effect as compared with the control, and that its antioxidant effect is 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 dd strain mice aged 7 weeks
An oral toxicity test of α-D-glucopyranosyl rutin product showed an acute toxicity test of 5 g / kg body weight.
No deaths were seen. Therefore, the toxicity of this substance is extremely low.

Further, when this test was carried out using high-purity 4 ^G- α-D-glucopyranosyl rutin powder prepared by the method of Reference Example A-4 described later, the same result was obtained.
The toxicity was found to be extremely low.

Hereinafter, as a reference example of the present invention, 4 ^G -α-
In Reference Example A, a production example of D-glucopyranosyl rutin,
Reference Example B describes an application example of 4 ^G- α-D-glucopyranosyl rutin to various compositions.

[0096]

[Reference Example A-1] <4 ^G- α-D-glucopyranosyl
Rutin> 1 part by weight of rutin is heated and suspended in 5 parts by weight of 50 V / V% ethanol aqueous solution, and separately dextrin (DE8)
8 parts by weight of water was dissolved in 45 parts by weight of water, and then these solutions were mixed with which Bacillus stearothermoph
(Ilus) -derived cyclomaltodextrin glucanotransferase (sold by Hayashibara Biochemical Research Institute, Inc.)
10 units were added per dextrin gum, the pH was adjusted to 6.0, and the reaction was carried out at 50 ° C. for 48 hours. When the reaction solution was analyzed by thin layer chromatography, most of it was converted into 4 ^G- α-D-glucopyranosyl rutin having glucose residues bonded in equimolar amounts or more.

The reaction solution was concentrated under reduced pressure to remove ethanol.
After that, inactivate the enzyme by heating and adjust to pH 5.0.
Glucoamylase (EC 3.2.1.3, biochemistry
Sold by Kogyo Co., Ltd.) 100 units per gram of solid
Then, the mixture was reacted at 50 ° C. for 5 hours. In addition, add this reaction solution.
Heat to deactivate the enzyme, filter, and filter the filtrate into an ion-exchange resin.
(H type and OH type) desalted, purified, concentrated, and solid
About 10 w / w% 4 ^G-Α-D-glucopyranosyl
  Small amount of rutin and large amount of glucose with rutin
Syrup-like products containing
The yield was about 95%.

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

Since this product contains a large amount of glucose, it can be used as a sweetener.

[0100]

[Reference Example A-2] <4 ^G- 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 dry-heat sterilized and aseptically added at the time of inoculation) and 500 parts by weight of a liquid medium containing Mucor Javanicus IFO4570
Was cultivated with shaking at a temperature of 30 ° C. for 44 hours. After completion of the culture, the mycelium was collected, and 0.5 parts were added to 48 parts by weight of the wet mycelium.
4M urea solution 50 dissolved in M acetate buffer (pH 5.3)
0 part by weight was added, and 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 of 0.
After 9-saturation, the mixture was left overnight at 4 ° C., and the salted-out product formed 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 prepared.

(2) 4 ^G- 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 by heating, and the pH was adjusted to 6.5 with 5N hydrochloric acid solution. Separately, 20 parts by weight of dextrin (DE30) was added to 10 parts by weight of water. It is dissolved by heating, and then these solutions are mixed to give a suspension-like solution containing a high amount of rutin, to which 10 parts by weight of the α-glucosidase preparation prepared by the method (1) was added, and p
The reaction was carried out at 55 ° C. for 40 hours while maintaining H6.5 while stirring.

In the same manner as in Reference Example A-1, the enzyme in the reaction solution was inactivated by heating, the pH was adjusted to 5.0, and glucoamylase was allowed to act on it. Further, the reaction solution was heated to inactivate the enzyme, filtered, and the filtrate was passed through a column of porous synthetic adsorbent resin, trade name Diaion HP-10 (sold by Mitsubishi Kasei Kogyo Co., Ltd.) by SV2. . As a result, a large amount of 4 ^G- 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. were discharged without being adsorbed. Then
After passing through the column with water and washing, an aqueous methanol solution was passed through to elute and collect 4 ^G- α-D-glucopyranosyl rutin and rutin, concentrate under reduced pressure, and pulverize to give a purity of about 7%.
A powder product containing 5 w / w% of 4 ^G- α-D-glucopyranosyl rutin and other rutin was obtained in a yield of about 90% based on the weight of rutin as a raw material.

This product is used as a highly water-soluble vitamin P enhancer, and also as a highly safe natural type yellow colorant, antioxidant, stabilizer, quality improver, preventive agent, therapeutic agent, ultraviolet absorber. As an anti-deterioration agent, etc., it can be advantageously used for various compositions such as food and drink, favorite food, feed, feed, anti-susceptive disease agent, cosmetics and plastic products.

[0104]

[Reference Example A-3] <4 ^G- α-D-glucopyranosyl
Rutin> 1 part by weight of rutin and dextrin (DE1
2) 10 parts by weight was mixed with 98 parts by weight of hot water at 80 ° C. to form a suspension-rich liquid containing rutin, to which 20 units of cyclomaltodextrin glucanotransferase was added per dextrin glam, pH 6.0, 75 ° C. The reaction was carried out for 64 hours while maintaining the above temperature and stirring. When the reaction solution was analyzed by thin layer chromatography, most of it was converted to α-glycosyl rutin having glucose residues bonded in equimolar amounts or more. The reaction solution was inactivated by heating, adjusted to pH 5.0, and reacted with glucoamylase in the same manner as in Reference Example A-1.

The reaction solution was deactivated by heating, filtered, and the filtrate was passed through a column of porous synthetic adsorbent, trade name Diaion HP-20 (sold by Mitsubishi Kasei Kogyo Co., Ltd.) at SV1.5. As a result, 4 ^G- α-D-glucopyranosyl rutin and unreacted rutin in the solution were adsorbed by the porous synthetic adsorbent, and glucose, salts, and the like flowed out without being adsorbed. Then, the column is flushed with water and washed, and then an aqueous solution of methanol is flushed through to wash the column with 4 ^G- α-D-glucopyranosyl.
Rutin and rutin were eluted, collected, concentrated under reduced pressure, and pulverized. This powder is heated and dissolved in hot water to a concentration of about 50 w / w%, and a small amount of rutin crystals is added as seed crystals to the crystals to agitate while stirring, to remove crystalline rutin by centrifugation and to concentrate the supernatant. , Powdered, 4 ^{G in the} form of powder with a purity of about 95 w / w%
An -α-D-glucopyranosyl rutin product was obtained in a yield of about 90% based on the weight of rutin of the raw material per solid matter.

Similar to Reference Example A-2, 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 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, favorite foods, feeds, feeds, anti-susceptive disease 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
− Put the glucopyranosyl rutin product in a glass container.
Dissolved in methanol at 25 w / w% with heating,
When kept at room temperature for 3 days, crystals precipitated on its inner wall.
It was This crystal was similarly prepared 4 ^G-Α-D-gluco
Pyranosyl rutin product concentration about 30 w / w% methanol
Solution as seed crystals and slowly stirring for 2 days
For a while, crystallize the resulting musket, and add a small amount to the crystals.
Crystallinity obtained by spraying with methanol and washing 4 ^G
-Allow α-D-glucopyranosyl rutin at 70 ° C overnight.
4 after air-drying with a purity of about 99w / w%^G-Α-D-gluco
Pyranosyl rutin powder was obtained.

This product is a high-purity 4 ^G- α-D-glucopyranosyl rutin, which is not only a highly water-soluble vitamin P enhancer but also a highly safe natural type yellow colorant and antioxidant. , Stabilizers, quality improvers, preventives, therapeutics, UV absorbers, deterioration inhibitors, etc., for use in various compositions such as food and drink, favorite foods, feeds, feeds, anti-susceptive disease agents, cosmetics, plastic products, etc. Can be used to advantage.

An example of a crystal photograph during crystallization from a methanol solution in this reference example is shown in FIG.

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

[0111]

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

The reaction solution was adjusted to pH 5.0 by deactivating the enzyme in the same manner as in Reference Example A-1, and reacted with glucoamylase. This reaction solution is deactivated by heating, filtered, and the filtrate is a porous synthetic adsorbent, trade name Amberlite XAD-
SV on column 7 (manufactured by Rohm & Haas)
The solution was passed at 1.5.

As a result, 4 ^G- 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 passing through this column with water and washing, the solution was passed through the column while increasing the concentration of the aqueous ethanol solution stepwise, and 4 ^G
A fraction containing a high amount of -α-D-glucopyranosyl rutin was collected, concentrated and pulverized to obtain a powdery 4 ^G- α-D-glucopyranosyl rutin product having a purity of about 93 w / w% based on the weight of rutin as a raw material. The yield was about 85%.

As in Reference Example A-2, 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 an improving agent, preventive agent, therapeutic agent, ultraviolet absorber, deterioration inhibitor, etc., it can be advantageously used for various compositions such as food and drink, favorite food, feed, feed, anti-susceptive disease agent, cosmetics and plastic products. .

[0116]

[Reference Example A-6] <High-purity ^4G- α-D-glucopyranosylrutin> ^4G- α-D obtained by the method of Reference Example A-5
-The 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 form 0.5 w / w. %, Slowly stirring 2
The crystals are eutectic for a day, the obtained musket is separated, and the crystals are washed by spraying a small amount of an aqueous solution of ethanol, and the obtained crystalline ^4G- α-D-glucopyranosyl rutin is heated at 80 ° C.
Vacuum dried overnight at 4 ^G- α-D with a purity of about 98 w / w%
-Glucopyranosyl rutin powder was obtained.

This product is high-purity 4 ^G- α-D-glucopyranosyl rutin, which is not only a highly water-soluble vitamin P enhancer but also a highly safe natural type yellow colorant and antioxidant. , Stabilizers, quality improvers, preventives, therapeutics, UV absorbers, deterioration inhibitors, etc., for use in various compositions such as food and drink, favorite foods, feeds, feeds, anti-susceptive disease agents, cosmetics, plastic products, etc. Can be used to advantage.

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

Further, when this crystal was examined by a powder X-ray diffractometer in the same manner as in Reference Example A-4, the main diffraction angles (2
θ) is 6.8 °, 8.1 °, 15.4 °, 17.8 °
And is different from the crystals crystallized from methanol obtained in Reference Example A-2.

Further, this crystal easily loses these diffraction angles by heating and drying to become an 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 (sold by Hayashibara Shoji Co., Ltd., registered trademark Mabit) 1,5
00 parts by weight are heated and concentrated under reduced pressure to a water content of about 2% or less, to which 15 parts by weight of citric acid and Reference Example A-
1 part by weight of a powdery 4 ^G- α-D-glucopyranosyl rutin product obtained by the method of 3 and a small amount of lemon flavor were mixed,
Then, it was molded and packaged according to a conventional method to obtain a hard candy.

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

[0123]

[Reference Example B-2] <Boiled butterbur> Peel the fluff, cut it to an appropriate length, and soak it in a thin saline solution for several hours. The powder obtained by the method of Reference Example A-5 It was boiled with a liquid containing a green colorant prepared by blending an α-glycosyl rutin product and Blue No. 1 to obtain a boiled green fluffy boiled water.

This product adds color as a material for various Japanese-style dishes and also exerts a physiological effect as a dietary fiber.

[0125]

[Reference Example B-3] <Fertilization> 1 part by weight of waxy starch was mixed with 1.
Mix 2 parts by weight and heat to gelatinize while adding 1.5 sugar.
Parts by weight, crystalline β-maltose (produced by Hayashibara Co., Ltd., registered trademark Sanmalto) 0.7 parts by weight, starch syrup 0.3 parts by weight and powdery 4 ^G -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 is a fertilizer with good flavor and mouthfeel,
It is a Japanese sweets of the kibi-dango style.

[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 4 ^G- α-D-glucopyranosyl syrup obtained by the method of Reference Example A-1 1 part by weight of the rutin product was mixed to obtain a mixed sweetener.

This 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 (produced by Hayashibara Co., Ltd., registered trademark FINETOSE)
1,200 parts by weight, shortening 1,000 parts by weight,
10 parts by weight of a powdery 4 ^G- 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 were mixed by an ordinary method to obtain a sand. A cream was produced.

This product is a vitamin P-fortified, yellow-colored sand cream that prevents the oxidation of fats and oils, and has a good mouthfeel, melting quality and flavor.

[0131]

[Reference Example B-6] <Orange juice> Orange juice 5
0 parts by weight, citric acid 0.1 parts by weight, sugar 5 parts by weight, high-purity 4 ^G- α-D-glucopyranosyl rutin powder 0.1 parts by weight obtained by the method of Reference Example A-6, L-ascorbic acid ( Vitamin C) 0.1 part by weight, an appropriate amount of perfume and 46 parts by weight of water were mixed, and then filled in a container and sterilized by an ordinary 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
4 ^G- α-D-glucopyranosyl powder obtained by the method
5 parts by weight of a rutin product and 40 parts by weight of ethanol were mixed with 26 parts by weight of purified water and appropriate amounts of a coloring agent and a fragrance to produce a bath agent.

The product is suitable as a skin beautifying agent and a skin whitening agent, and may be diluted 100 to 10,000 times in hot water for bathing before use. This product is the same as for hot water for bathing,
It can also be advantageously carried out by diluting with face-washing water, lotion and the like.

[0135]

[Reference Example B-8] <Emulsion> 0.5 part 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. , Behenyl alcohol 0.5 part by weight, avocado oil 1 part by weight, high-purity 4 ^G- α-D-glucopyranosyl rutin powder 1 part by weight obtained by the method of Reference Example A-4, vitamin E and an appropriate amount of preservative were always added. The mixture was heated and dissolved according to the method, 1 part by weight of L-sodium lactate, 5 parts by weight of 1,3-butylene glycol, 0.1 part by weight of carboxyvinylponimer and 85.3 parts by weight of purified water were added, and the mixture was homogenized and emulsified. ,
Further, an appropriate amount of fragrance was added and mixed with stirring to produce an emulsion.

This product has antioxidant properties and is highly stable and can be advantageously used as a high-quality sunscreen, skin-refining agent, skin-whitening agent and the like.

[0137]

Reference Example B-9 <Cream> Polyoxyethylene glycol monostearate 2 parts by weight, self-emulsifying glyceryl monostearate 5 parts by weight, high purity 4 ^G- 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 were dissolved by heating according to a conventional method, and 2 parts by weight of L-lactic acid and 1,3-butylene glycol 5 were added thereto. By weight, 66 parts by weight of purified water was added, the mixture was homogenized and emulsified, and then an appropriate amount of flavor was added and mixed with stirring to produce a cream.

This product has antioxidative properties and high stability,
It can be advantageously used as a high-quality sunscreen, skin-refining agent, and skin-whitening agent.

[0139]

Reference Example B-10 <Antioxidant> 10 parts by weight of powdered 4 ^G- 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 blended with oily foods such as margarine and butter cream, unsaturated fatty acids, oily vitamins, anti-sensitivity agents such as oily hormones, cosmetics such as emulsions and creams in an amount of about 0.01 to 5.0 W / W%. Therefore, 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 high-purity 4 ^G- 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 blended with oily foods such as margarine and butter cream, unsaturated fatty acids, oily vitamins, anti-sensitivity agents such as oily hormones, cosmetics such as emulsions and creams in an amount of about 0.01 to 5.0 W / W%. In addition, about 0.01 to 2.0 W / W% is added to foods and drinks containing natural pigments that easily discolor, which is advantageous as an antioxidant, stabilizer, anti-fading agent, quality improving agent, etc. Available.

Examples of the present invention will be described below.

[0144]

Example 1 <Tablet> 20 parts by weight of ascorbic acid, 13 parts by weight of crystalline β-maltose, 4 parts by weight of corn starch and 3 parts by weight of the powdery α-glucosylrutin product obtained by the method of Reference Example A-2 were homogeneous. Diameter of 12m after mixing to
Tablets were obtained by using a m, 20R punch.

This product is ascorbic acid and 4 ^G- α-D-
It is a complex vitamin preparation with glucopyranosyl rutin. It has good stability of ascorbic acid and is easy to take.

[0146]

[Example 2] <Capsule> Calcium acetate / monohydrate 1
0 parts by weight, L-magnesium lactate / trihydrate 50 parts by weight,
57 parts by weight of maltose, 20 parts by weight of a powdery 4 ^G- 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 added. After mixing and granulating with a granule molding machine, they were encapsulated in gelatin capsules according to a conventional method to produce capsules each containing 150 mg.

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

[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 the 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 1 part by weight of α-D-glucopyranosyl rutin powder and 0.5 part by weight of peppermint oil, and further uniformly mixing.

This product has antioxidative properties and high stability,
It can be advantageously used as a high-quality sunscreen, skin-refining agent, skin-whitening agent, etc., and also as an accelerator for healing 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, and purified and filtered according to a conventional method to be pyrogen-free. The solution is placed in a 20 mL ampoule and 4 ^G- α-D-
Glucopyranosyl rutin was dispensed to 200 mg, dried under reduced pressure, and sealed to prepare an injection.

This injection can be administered intramuscularly or intravenously as a single substance or as a mixture with other vitamins and minerals. Further, this product does not need to be stored at a low temperature, and its solubility in physiological saline and the like at the time of use is extremely good.

This product not only supplements vitamin P, but also exerts effects such as removal of active oxygen and suppression of production of lipid peroxide as an antioxidant. Viral diseases, bacterial diseases, cardiovascular diseases, and malignant diseases. It can be advantageously used as a preventive 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, and Purify and pyrogen-free according to the method and place this solution in a sterile plastic container.
An injection was prepared by filling 0 mL each.

This product is an injection for not only vitamin P supplementation but also calorie supplementation, mineral supplementation, further removal of active oxygen and suppression of lipid peroxide production.
Promotion of treatment after illness, promotion of recovery, 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] <Nutrient for tube feeding> Crystalline α-maltose 20
Parts by weight, glycine 1.1 parts by weight, sodium glutamate 0.18 parts by weight, salt 1.2 parts by weight, citric acid 1 part by weight, calcium lactate 0.4 parts by weight, magnesium carbonate 0.1 parts by weight, Reference Example A -4 ^G powder obtained by the method of No. 5
0.1 parts by weight of α-D-glucopyranosyl rutin product,
A formulation consisting of 0.01 parts by weight thiamine and 0.01 parts by weight riboflavin is prepared. Each 24 g of this mixture was filled in a laminated aluminum pouch and heat-sealed to prepare a tube feeding agent.

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

[0157]

Industrial Applicability As described in the text, the novel substance 4 ^G- α-D-glucopyranosyl rutin of the present invention increases the water solubility to 30,000 times or more while maintaining the molecular extinction coefficient (yellowness) of rutin. Moreover, it is substantially tasteless and odorless, has no fear of toxicity, and is easily hydrolyzed into rutin and D-glucose in the body to exert rutin's original physiological activity.

Further, since this 4 ^G- 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 economical. It has excellent properties, and its industrial implementation is easy.

It was also found that the reaction concentration of rutin can be increased and ^4G- α-D-glucopyranosyl rutin can be easily produced at a high concentration, and at the same time, in the purification of this reaction solution, 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, 4 ^G -α-D-glucopyranosyl
By establishing a method for producing 4 ^G- α-D-glucopyranosyl rutin with higher purity by crystallizing and separating it from a supersaturated solution of rutin, its industrial realization becomes extremely easy.

Further, 4 ^G -α-obtained in this way
D-glucopyranosyl rutin is not only a highly safe natural vitamin P enhancer but also a yellow colorant,
Antioxidants, stabilizers, quality improvers, preventive agents, therapeutic agents, ultraviolet absorbers, deterioration inhibitors, etc.
It is advantageously used for application to various compositions such as feeds, baits, anti-susceptive diseases agents, skin-beautifying agents, skin-whitening agents, and plastic products.

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

[Brief description of drawings]

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

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

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

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

FIG. 5 is a crystal photograph of the present invention's 4 ^G- α-D-glucopyranosyl rutin crystallized (40 times magnification).

─────────────────────────────────────────────────── ─── Continued Front Page (51) Int.Cl. ⁷ Identification Code FI A61P 29/00 101 A61P 29/00 101 31/04 31/04 31/18 31/18 35/00 35/00 37/00 37 / 00 // C07H 17/07 C07H 17/07 (58) Fields surveyed (Int.Cl. ⁷ , DB name) A61K 31/7048 C07H 17/07 REGISTRY (STN) CA (STN)

Claims (4)

(57) [Claims] 1. 4 ^G -α-D- represented by the following chemical formula 1.
The 4 ^G- α containing glucopyranosyl rutin
-D-glucopyranosyl rutin is produced by an in vivo enzyme.
Prevent or cure by rutin produced by hydrolysis.
Anti-susceptive agent for the disease to be treated . [Chemical 1] 2. The anti-susceptive disease agent according to claim 1, wherein 4 ^G- 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. With 4. 4 ^{G-.alpha.-D-glucopyranosyl} rutin, thiamine, riboflavin, according to either the claims 1 to 3, characterized in that it contains vitamins chosen from vitamin C and vitamin E Anti-susceptive disease agent.