JP6155632B2 - Isoflavone conversion promoting composition - Google Patents

Description

  The present invention is used in foods and drinks, feeds, pharmaceuticals, etc., and when taken orally, osteoporosis prevention / treatment by improving bone density in the intestine, breast cancer prevention / treatment, cholesterol accumulation suppression effect, immune enhancement effect, etc. The present invention relates to a composition that promotes genistein, daidzein, and equol production.

  It is known that isoflavones are abundant in soybeans. Soy isoflavone is one secondary metabolite formed during growth, with a content of about 0.25% of soy. It mainly contains free genistein, daidzein, glycitein and conjugated genistin, daidzine and the like. Many studies have shown that soy isoflavones have a wide range of functions such as anti-cancer, osteoporosis prevention, cardiovascular disease prevention, women's menopause improvement, renal function improvement, immunity improvement, weight gain, etc. ing. Among these, with recent rapid aging, osteoporosis is becoming a major problem because bone density decreases with age and bone fractures easily occur.

  In order to prevent osteoporosis, in order to increase bone mass from a young age, it is important to consume a large amount of calcium-rich dairy products and fish, etc., from moderate exercise and daily use. In addition, aldonic acids such as vitamin K, lactobionic acid, oligosaccharides and minerals, casein phosphopeptide, soybean isoflavone, resistant oligosaccharides, chondroitin and isoflavone, carotenoids and flavonoids derived from Wenzhou mandarin orange (and derivatives thereof) ), Or β-glucooligosaccharide, soybean isoflavone, vitamin D, or the like is also effective as a so-called health food.

  In particular, genistein, daidzein and equol, which are metabolites of soybean isoflavones, are isoflavones with high antioxidant capacity and strong estrogenic activity, and have already been reported to be effective as therapeutic agents for osteoporosis (Patent Literature). 1).

  Furthermore, as the action of these isoflavones, plasma cholesterol lowering action, breast cancer, prostate cancer suppression and the like have been reported. (Non-patent document 1) Furthermore, anticancer effects of genistein and daidzein have been reported (non-patent document 2). On the other hand, since the metabolism from daidzein to equol is mainly carried out by intestinal bacteria, it is known that the production amount varies depending on the individual intestinal bacterial flora, male and female differences and race differences. In fact, about 60 to 70% of Westerners are reported to have low equol production capacity. Therefore, DFAIII (difructose-dianhydride III) (Non-patent document 3) and fructooligosaccharide (Non-patent document 4) as well as substances that promote the production of equol, It is said that it is effective for conversion.

JP 2005-232074 A

“Chemistry and Biology”, published by the Japanese Society for Agricultural Chemistry, Vol. 44, no. 3, p. 151-153 (2006) Ichizo Yagasaki, Masaki Takatsuki, Yutaka Miura, “Soy Protein Research”, published by Fuji Protein Research Foundation, Vol. 16, p122-127 (2003) The 49th Annual Meeting of the Japanese Agricultural Chemical Society, published by the Japanese Agricultural Chemical Society, p. 97 (2005) Abstracts of the 56th Annual Meeting of the Japanese Society of Nutrition and Food, published by the Japanese Society of Nutrition and Food, p. 138 (2002)

  However, fructooligosaccharides, DFAIII, and lactobionic acid increase the production of equol, but their effects are insufficient. Although β gluco-oligosaccharide has been reported, it focuses on the enhancement of Ca absorption and is not intended for the conversion of isoflavones.

  In addition, the intake of microorganisms capable of converting to equol is still a technical issue in order to reach the large intestine alive, and there is a need for inexpensive pharmaceuticals and food materials that can effectively promote the production of equol. Development is required.

  In addition, equol is not contained in food, and there is no way to consume equol at a low cost. Therefore, the development of food materials that can significantly promote the production of equol has been desired.

  Accordingly, an object of the present invention is to provide a composition capable of significantly promoting the production of genistein, daidzein and equol.

  As a result of intensive studies to solve such problems, the present inventors have found that a composition containing cellooligosaccharide or derivatives thereof having 2 to 10 β1,4-linked glucose as an active ingredient, preferably cellobiose is an active ingredient. As a result, it was found that the production of genistein, daidzein, and equol was promoted by ingesting the composition contained therein, and the present invention was completed.

  That is, the present invention provides an equol production-promoting composition comprising cellooligosaccharide or a derivative thereof as an active ingredient, preferably genistein, wherein cellooligosaccharide or a derivative thereof has cellobiose as a main component. , Daidzein and equol production promoting composition.

  In addition, the present invention is characterized by containing the above-mentioned genistein, daidzein and equol production promoting composition, 1) an osteoporosis prevention and / or treatment agent, and 2) containing the above equol production promotion composition. Food / beverage products that increase bone density, prevent breast cancer / prostate cancer, reduce cholesterol, or enhance immunity 3) Increase bone density Feed or pet food, 4) breast cancer / prostate cancer preventive agent, 5) cholesterol lowering agent.

  The composition for promoting genistein, daidzein and equol production of the present invention has an extremely high effect of promoting equol production. Therefore, when combined with pharmaceuticals and foods and drinks, it prevents vascular diseases due to cholesterol lowering action, breast cancer, prostate cancer prevention, and osteoporosis. Effect of prevention and / or treatment of symptom. In addition, when blended in feed or pet food, bone density is enhanced, and effects such as preventing leg weakness in pigs, strengthening eggshells of laying hens, and preventing osteoporosis in dogs and the like are exhibited.

It is a graph which shows a time-dependent change of the amount of genistin. It is a graph which shows a time-dependent change of the amount of daidzins. It is a graph which shows the production amount of genistein. It is a graph which shows the production amount of daidzein. It is a graph which shows a time-dependent change of the amount of genistein. It is a graph which shows a time-dependent change of daidzein. It is a graph which shows the production amount of an equol amount.

  Hereinafter, the present invention will be described in detail.

  In the present invention, the cellooligosaccharide is a product in which 2 to 10 glucoses are linked by β1,4 bonds. Among these, cellobiose and cellotriose are preferable as active ingredients.

  Cellulo-oligosaccharides in this invention include cellulase using cellulose as a substrate, partial decomposition of cellulose with mineral acids such as hydrochloric acid and sulfuric acid, partial decomposition with heated hot water, hydrolase, phosphorolytic enzyme Any of the methods such as reverse polymerization reaction by the method may be used.

  The active ingredient prepared by the above method can be used as a composition for promoting genistein, daidzein and equol production by itself or together with a solid or liquid excipient known in the art.

  Cellooligosaccharides can also be derivatized in a timely manner according to the usage form and purpose. Derivatives may be esterified, phosphorylated, sulfonated, glucosylated, or in the form of their salts.

  Examples of solid excipients include lactose, sucrose, glucose, sugar alcohols, corn starch, gelatin, dextrin, starch, oligosaccharides, indigestible dextrin, indigestible starch, and the like. Examples of the liquid excipient include water, glycerin, fatty oil, sorbitol and the like.

  The composition for promoting genistein, daidzein and equol production of the present invention is suitable for timely use, for example, an antibacterial agent such as berberine chloride, agate powder, magnesium oxide, an antibacterial agent such as oxytetracycline or bicozamycin, such as sodium propionate. It may contain other agents such as fungicides, e.g. Praziquantel, anthelmintic agents such as ivermectin and / or antioxidants, pigments, flavorings, flavorings, conventional additives such as enzymes, Since production of genistein, daidzein and equol is further promoted, isoflavones, soybean extract and the like may be added in advance. In addition, it may be added to general soybean foods (soy milk, natto, tofu, ganmodoki, miso, soy sauce, kinako, etc.) and formulated into powders, granules, liquids, tablets, etc. by conventional methods can do. Furthermore, lactic acid bacteria, bifidobacteria, yeast, Bacillus subtilis, butyric acid bacteria and / or bacilli may be added.

  These compositions can be administered per se to humans or animals and can be used as pharmaceuticals, food and drink, feed or pet food. When used as a food or drink or feed, it is convenient and preferable to add it to the material in advance.

  The food, beverage, feed or pet food to which the genistein, daidzein and equol production promoting composition of the present invention should be added is not particularly limited as long as it is generally used. Examples thereof include foods and drinks, feeds and pet foods made from soybeans, egg whites, milk, flour, seafood and the like.

  Examples of such foods and drinks include, for example, health foods such as tablets, powders, granules, syrups and capsules, processed foods such as breads, confectionery, cookies, biscuits and chocolate, milk, yogurt, Examples include dairy products such as ice cream, carbonated drinks, soft drinks, soft drinks with fruit juice, drinks with fruit juice, drinks such as medicinal drinks, side dishes and processed foods using soybeans, fish meat, livestock meat, and the like.

  Examples of feed and pet food include corn, rice, wheat, milo, soybean meal, bran, beet pulp, cellulose, defatted rice bran, fish meal, defatted milk powder, dried whey, fats and oils, alfalfa meal, North Sea meal, soybean fats and oils, Powdered beef tallow, wheat flour, rapeseed oil, meat and bone meal (feather meal), animal fat, calcium phosphate, corn gluten meal, molasses, corn germ meal, calcium carbonate, tricalcium phosphate, sodium chloride, choline chloride, vitamins (vitamins A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin D, vitamin E, calcium pantothenate, nicotinamide, folic acid, etc.), amino acids (lysine, methionine, etc.), trace inorganic salts (magnesium sulfate, iron sulfate) , Copper sulfate, zinc sulfate, iodide Potassium, cobalt sulfate, etc.), can be mentioned those prepared by mixing appropriate and viable cell agents.

  Moreover, as a pharmaceutical, it can be ingested in the form of powders, tablets, granules, capsules, suspensions, syrups, liquids for internal use, troches and the like.

  The content of the genistein, daidzein, and equol production promoting composition of the present invention depends on the type and weight of the target human or animal, but when added to foods and drinks, feed or pet food, The sugar is generally 0.01 to 10.0% by weight, preferably 0.01 to 5.0% by weight. Moreover, when using as a pharmaceutical, it is preferable to design a formulation so that 10 mg to 100 g, preferably 500 mg to 10 g, more preferably 1 g to 5 g can be ingested as cellooligosaccharide per 50 kg adult per day.

EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated further in detail, this invention is not limited to these Examples.

Example 1
[Confirmation effect of genistein, daidzein and equol production by rats]
1. AIN93G 20% casein feed as a sample basic feed (casein 200 g, dextrin 649.486 g, corn oil 50 g, cellulose 50 g, AAIN-93G mineral mix 35 g, AIN-93 vitamin mix 10 g, L-cystine 3 g, choline bitartrate 2 0.5 g, t-butylhydroquinone 0.014 g) was used as a 3 wt% mixed powder bait in which cellooligosaccharide (trade name: Sancellobio K manufactured by Nippon Paper Industries Co., Ltd.) was mixed to 3.0 wt%. Further, as a comparative control, fructooligosaccharide (FOS) (manufactured by Wako Pure Chemical Industries, Ltd.) 3 wt% mixed powder bait was used. Genistin, daidzin, genistein, daidzein, and equol were manufactured by Wako Pure Chemical Industries.
2. Laboratory animals, rearing

6-week-old SD rat male rats (6-week-old, Japan SLD) were grouped in 6 rats, and acclimated breeding was carried out for 1 week with free intake of powder diet CRF-1 and water. At the end of acclimatized breeding, the 0th week was set, and from the 0th week, the AIN93G 20% casein diet was freely consumed in the cellooligosaccharide administration and fructooligosaccharide administration prepared above. The control group was fed with AIN93G 20% casein diet.

3. Two weeks after adjustment of the cecal microorganisms, the cecum of each group is removed and the cecal contents are collected. PB (phosphate buffer; NaH2PO4 · 2H2O 50 mmol / L, Na2HPO4 · 12H2O 50 mmol / L, L-cysteine 0.5 g / L, Na2CO3 3 g / L, resazurin sodium salt (0.1%) 1 ml / L, pH 6.5), add carbon dioxide, stir, and make a 10-fold dilution of cecal contents.

4). Metabolites measurement after adding genistin, daidzin, genicytein, daisein basic diet (AIN93G 20% casein diet) diet, 3% sanserobio diet, 3% fructooligo diet Dispense 2 ml each into a vial (5 ml volume), and adjust the added sample so that genistin, daidzin, genicytein and daisein have a final concentration of 1 mmol / L. Carbon dioxide is enclosed in each sample and incubated at 37 ° C. For genistin and daidzin, 100 μL of a sample is collected 30 and 60 minutes after the start of incubation, and the reaction is stopped by adding the same amount of methanol. For genisitein and dycein, 100 μL of sample is collected 24 hours and 48 hours after the start of incubation, and the same amount of methanol is added to stop the reaction. In addition, 100 microliters is extract | collected from each vial bottle as a sample at the time of 0 minutes before incubation, and it preserve | saves at -80 degreeC.

5. The analytical analyzer was a Waters UPLC / MC / MC system (LC is ACQUITY UPLC, MS / MS is ACQUITY TQD tandem quadrupole mass spectrometer). Analytical conditions were ACQUITY UPLC BEH C-18 1.7μm, 2.1 x 50mm column (Waters), column temperature 40 ° C, mobile phase A 1% formic acid aqueous solution, mobile phase B 1% formic acid acetonitrile solution, flow rate The gradient conditions were 0.6 mL / min, 0-0.3 min: 97% A solution, 1.3-3 min: 10% A solution, 3.2-5 min: 97% A solution. The ionization method used ESI positive mode.

5-1. Experimental Results Aglyconization of isoflavone glycosides Figure 1 shows the results of changes in the amount of genistin over time, Figure 2 shows the results of changes in the amount of daidzine, and Figure 3 shows the amount of genistein produced. Is shown in FIG. As a result, in the cellooligosaccharide administration group, genistin decreased significantly faster than other groups, and in parallel, the amount of genistein, an aglycone of genistin, increased. Similarly, in the cellooligosaccharide administration group, it was found that daidin decreased significantly faster than other groups, and in parallel, the amount of daidcein, which is an aglycon of daidin, increased. This is thought to be because β-glucosidase activity of enterobacteria increased by cellooligosaccharide administration, and as a result, aglyconization of isoflavone glycosides was promoted.

5-2. Experimental results Time-dependent changes in the amounts of genistin and daidzein and time-dependent changes in the amount of equol in each group are shown in FIG. 5, the results of time-dependent changes in the amount of genistein are shown in FIG. . It was found that the amount of genistein decreased less in the cellooligosaccharide administration group than in the control and fructooligosaccharide administration groups. Therefore, it was suggested that the absorbed amount of genistein, which is the active substance, may be reduced. Moreover, although the difference in the amount of daidzein was not observed in each group, it was found that the production amount of equol was higher in the cellooligo administration group than in the other groups in 24 hours of incubation. Because daidzein is known to be converted to equol via dihydrodaidzein, the cellooligosaccharide administration group activates bacteria that have an enzyme that converts dihydrodaidzein to equol, or degrades equol. The bacterial group may be less than the control and fructooligosaccharide-administered group.

Claims (2)

  A cellooligosaccharide having a degree of polymerization of 2 to 10, or an esterified derivative thereof, a phosphorylated derivative thereof, a sulfonated derivative thereof, a glycosylated derivative thereof, and the cellooligosaccharide derivative selected from at least one Genistein, daidzein production promoting composition.   The composition for promoting genistein and daidzein production according to claim 1, wherein the cellooligosaccharide is cellobiose.

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