JP2006193452A - Composition for inhibiting osteoclast differentiation or proliferation, food and beverage containing the composition for inhibiting osteoclast differentiation or proliferation - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a composition for inhibiting osteoclast differentiation or proliferation, which is expected to have sufficient prevention or treatment and amelioration effect on osteoporosis by inhibiting osteoclast differentiation or proliferation and controlling bone resorption and has high safety, and a food and beverage containing the composition for inhibiting osteoclast differentiation or proliferation. <P>SOLUTION: The composition for inhibiting osteoclast differentiation or proliferation comprises an extract of an enlarged part of Zizania latifolia and/or a ground enlarged part of Zizania latifolia as an active ingredient. The extract of an enlarged part of Zizania latifolia is obtained by extracting an enlarged part of Zizania latifolia with chloroform or ethanol. The food and beverage contains preferably 0.5-70% by mass of the composition for inhibiting osteoclast differentiation or proliferation calculated as dried enlarged part of Zizania latifolia. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  TECHNICAL FIELD The present invention relates to an osteoclast differentiation / proliferation inhibitor composition containing a makomotake extract and / or a pulverized mash compost as an active ingredient, and a food and drink containing the osteoclast differentiation / proliferation inhibitor composition.

  In the modern Japanese society where aging is progressing year by year, an increasing number of people suffer from various diseases associated with aging. For example, osteoporosis, which is one of the diseases associated with aging, refers to a condition in which the bone mass decreases and the bone becomes very brittle.

  In normal bone, bone resorption, in which osteoclasts break down old bone parts, and bone formation in which osteoblasts create new bone where bone resorption is performed, are balanced, resulting in a constant bone mass. It is kept. However, due to insufficient intake of calcium, a decrease in calcium absorption ability, disorder of hormone balance, etc., the balance of bone turnover is lost, and bone resorption exceeds bone formation, leading to osteoporosis. In particular, postmenopausal women are prone to osteoporosis due to hormonal balance disturbance.

  Therefore, for osteoporosis, it is considered necessary to suppress bone resorption or promote bone formation.

  For this reason, active vitamin D3 preparations, vitamin K2 preparations, estrogen preparations, calcitocin preparations, calcium preparations, bisphosphonate preparations, and the like are used as conventional therapeutic agents for osteoporosis. There was a problem of side effects.

  1) Active vitamin D3 preparation: Anorexia, malaise, renal failure, etc. due to excessive increase in blood calcium level.

  2) Vitamin K2 preparation: Decreased effect of a drug called warfarin taken in the case of myocardial infarction.

  3) Estrogen preparation: Increased risk of myocardial infarction and stroke, increased incidence of breast cancer and endometrial cancer.

  4) Calcitocin preparation: nausea, hot flush, etc.

  5) Calcium preparation: gastrointestinal disorders.

  6) Bisphosphonate formulation: digestive symptoms, especially esophageal ulcers.

  7) Synergistic effect of side effects by the combined use of active vitamin D3 preparation and calcium preparation.

  Therefore, development of a safer osteoporosis treatment agent that has no side-effect problems even when ingested for a long period of time has been promoted. For example, Patent Document 1 listed below discloses a basic protein fraction derived from milk as an active ingredient. A bone strengthening agent is disclosed.

  Patent Document 2 listed below discloses an osteoclast differentiation inhibitor production promoter containing milk-derived basic protein fraction, particularly lactoferrin as an active ingredient.

  Patent Document 3 listed below discloses a bone strengthening agent containing casein phosphopeptide and genistein as active ingredients.

  Patent Document 4 listed below discloses an osteoporosis preventive / therapeutic agent containing collagen or an enzyme degradation product of collagen as an active ingredient.

Patent Document 5 listed below discloses a composition for promoting bone formation and preventing bone mineral content reduction using isoflavone as a main active ingredient.
Japanese Patent No. 3112737 JP 2004-115509 A JP 2001-302539 A Japanese Patent Laid-Open No. 11-12192 Japanese Patent Laid-Open No. 10-114653

  Various osteoporosis therapeutic agents containing the ingredients derived from foods described in the above patent documents as active ingredients can be easily ingested as health foods, etc., and there are few side effects, but all can be expected to be sufficiently satisfactory. I could not say.

  Therefore, an object of the present invention is to inhibit osteoclast differentiation / proliferation and inhibit osteoclast differentiation / proliferation, thereby suppressing osteoclast differentiation / proliferation. It is providing the food-drinks containing the composition and this osteoclast differentiation / growth inhibitory composition.

  As a result of intensive studies to achieve the above object, the present inventors have found that a component that inhibits osteoclast differentiation / proliferation is contained in makomotake, and the present invention has been completed.

  That is, the osteoclast differentiation / proliferation inhibitory composition of the present invention is characterized by containing a makomotake extract and / or a pulverized mashroom as an active ingredient.

  In the osteoclast differentiation / proliferation-inhibiting composition of the present invention, it is preferable that the above-mentioned extract of Momotake is obtained by extracting Momotake with chloroform or ethanol.

  The composition for inhibiting osteoclast differentiation / proliferation according to the present invention has an excellent osteoclast differentiation / proliferation inhibitory effect by containing the makomotake extract and / or the crushed mashlet as an active ingredient. Expected to be effective in preventing and treating osteoporosis. Makomotake is a material with abundant food experience and has high safety.

  Moreover, the food / beverage products of this invention are characterized by containing the said osteoclast differentiation / growth inhibitory composition.

  In the food-drinks of this invention, it is preferable to contain the said osteoclast differentiation / growth inhibitory composition 0.5-70 mass% in conversion of dry makomotake.

  Since the food / beverage products of this invention contain the said osteoclast differentiation / proliferation inhibitory composition, it can be ingested easily over a long period of time, and the prevention / treatment improvement of osteoporosis can be expected.

  According to the present invention, osteoclast differentiation which contains a component derived from Makotake which has been confirmed to be safe by abundant dietary experience as an active ingredient, is highly safe and can be expected to have sufficient osteoporosis prevention / treatment improvement effect. A growth inhibitory composition can be provided.

  Moreover, by including the above-mentioned osteoclast differentiation / proliferation inhibitory composition in a food or drink, it is possible to provide a food or drink that can be easily ingested and can be expected to have an effect of preventing or treating osteoporosis.

  Makomotake is a Chinese vegetable grown from East Asia to Southeast Asia, mainly in China and Taiwan, with young stems of Macomo, which is an aquatic plant belonging to the genus Macomo and perennials, grown sproutingly with Aspergillus niger. In recent years, cultivation has been carried out in Japan, and these commercial products can be easily obtained.

  The makomotake extract used in the present invention is dried and pulverized if necessary, and then extracted at an arbitrary temperature using an organic solvent, water, etc., preferably 5 to 50 times the mass (mass ratio) of makomotake. Can be obtained. After extraction, the residue may be removed by centrifugation, filtration or the like, concentrated as necessary, or further dried.

  As the organic solvent, for example, hexane, chloroform, ethyl acetate, butanol, propanol, 2-propanol, ethanol, methanol and the like can be used. In the present invention, chloroform and ethanol are preferably used for food. Ethanol is particularly preferably used because it can be used safely.

  In the case of extraction with an organic solvent, it is preferable to sufficiently remove the organic solvent after extraction by concentration under reduced pressure, air drying, or the like. The extraction temperature and extraction time are not limited because they differ depending on the extraction solvent. For example, when chloroform is used, extraction is preferably performed at room temperature (20 ° C.) to 50 ° C. for 2 to 12 hours, using ethanol. If it is, it is preferably extracted at room temperature (20 ° C.) to 70 ° C. for 2 to 24 hours.

  The pulverized mushroom mash used in the present invention can be obtained by drying the momotake mushroom as necessary and then pulverizing it with a known pulverizing means (for example, a pulverizer, a homogenizer, a mill, a jet mill, a ball mill, etc.). There is no restriction | limiting in particular in the shape and magnitude | size of a ground material, According to a use, it can determine suitably.

  The composition for inhibiting osteoclast differentiation / proliferation of the present invention may use the above-mentioned extract of Momotake and / or the pulverized product of Momotake as it is, but isoflavone, collagen peptide, lactoferrin, casein phosphopeptide, fish bone calcium, shellfish calcium, etc. One or more of these known osteoporosis preventive / therapeutic components may be added, and in addition, inorganic salts, organic acids, carbohydrates, proteins, peptides, amino acids, lipids may be added. It can mix | blend suitably. By blending the above components, a synergistic effect with the extract of the mushroom and / or the mushroom mushroom can be expected.

  Examples of the intake form of the osteoclast differentiation / proliferation inhibiting composition of the present invention include liquids, powders, tablets, pills, fine granules, granules, capsules, jellies, chewables, pastes, etc. Insofar as possible, foods and pharmaceutically acceptable carriers, excipients, sugars, sweeteners, flavors, acidulants, colorants, and other auxiliary additives may be used.

  The effective intake amount of the composition for inhibiting osteoclast differentiation / proliferation of the present invention may be appropriately set depending on the body weight, sex, age, etc. of the intake person, but is usually 0.01-5 g per kg body weight in terms of dry makomotake. / Day, preferably 0.05 to 0.5 g / day.

  The osteoclast differentiation / proliferation-inhibiting composition of the present invention can also be ingested by being mixed with food and drink. For example, (1) soft drink, carbonated drink, fruit drink, vegetable juice, lactic acid bacteria drink, milk drink, soy milk , Mineral water, tea beverages, coffee beverages, sports beverages, alcoholic beverages, jelly beverages and other beverages, (2) tomato puree, canned mushrooms, dried vegetables, pickled vegetables, etc. (3) dried fruits, jams Processed fruits such as fruit puree and canned fruit, (4) spices such as curry powder, wasabi, ginger, spice blend, seasoning powder, (5) noodles such as pasta, udon, soba, ramen, macaroni (raw noodles, (Including dried noodles), (6) breads such as bread, confectionery bread, cooking bread, donuts, (7) alpha rice, oatmeal, rice cake, batter flour, etc. (8) baked goods, bis Candy, rice confectionery, candy, chocolate, chewing gum, snack confectionery, frozen confectionery, candied confectionery, Japanese confectionery, Western confectionery, half confectionery, pudding, ice cream confectionery, (9) red beans, tofu, natto, kinakome, yuba , Legume products such as boiled beans, peanuts, (10) processed foods of honey, royal jelly, (11) meat products such as ham, sausage, bacon, (12) yogurt, pudding, condensed milk, cheese, fermented milk, butter, ice cream, etc. Dairy products, (13) processed egg products, (14) processed fish such as dried fish, salmon, chikuwa, and fish sausage, processed seaweed such as dried seaweed, kelp, and boiled fish, and processed fish such as tarako, kazuko, salmon roe Egg, (15) Dashi-no-moto, soy sauce, vinegar, mirin, consomme base, Chinese base, concentrated soup stock, dressing, mayonnaise, ketchup Seasonings such as miso, edible oils and fats such as salad oil, sesame oil, linoleic oil, diacylglycerol, safflower oil, (16) soup (including powder and liquid), semi-cooked foods, side dishes, retort foods , Chilled foods, semi-cooked foods (for example, cooked rice, crab balls).

  The blending amount of the osteoclast differentiation / proliferation inhibiting composition in the food and drink is preferably 0.5 to 70% by mass, more preferably 1 to 20% by mass in terms of dry makomotake. If the blending amount is less than the above range, it is difficult to take a sufficient amount in one meal. If the blending amount is more than the above range, the taste and quality stability of the food and drink will be defective, and in beverages and processed foods , Because the original flavor of them is impaired.

  After 5,000 g of commercially available mushrooms were dried in the sun, the mixture was pulverized using a mixer to obtain 350 g of pulverized mushrooms.

  1,000 ml of chloroform was added to 100 g of the crushed bamboo shoot obtained in Example 1, and the mixture was extracted with stirring at room temperature for 2 hours. After removing the residue by suction filtration, the resulting extract was completely removed of chloroform using a rotary evaporator, and then dried under reduced pressure to obtain 4.0 g of Makotake extract.

  1,000 ml of ethanol was added to 100 g of the crushed bamboo shoot obtained in Example 1, and extraction was performed with stirring at 30 ° C. for 3 hours. After removing the residue by suction filtration, the obtained extract was concentrated under reduced pressure using a rotary evaporator to remove ethanol to obtain 21.0 g (87% solid content) of paste-like Makotake extract.

  1,000 ml of water was added to 100 g of the crushed bamboo shoot obtained in Example 1, and extraction was performed with stirring at 95 ° C. for 1 hour. After removing the residue by suction filtration, the obtained extract was concentrated under reduced pressure using a rotary evaporator to obtain 51.0 g (82% solid content) of paste-like Makotake extract.

<Test Example 1>
With respect to the extract of the mushroom obtained in Example 2, the osteoclast differentiation / proliferation inhibitory activity was examined.

  First, RAW264.7 cells (obtained from American Type Culture Collection, ATCC No. TIB71) transfected with estrogen receptor α: ERα were selectively cultured in a medium containing 600 μg / ml G418, and then 100 U / ml Using phenol red-free αMEM medium (GIBCO) containing penicillin, 100 μg / ml streptomycin, and 10% (v / v) heat-inactivated CD-FBS, 37 ° C., 5% carbon dioxide The cells were grown under the following conditions.

Next, the above RAW264.7 cells suspended in phenol red-free αMEM medium containing 10% (v / v) heat-inactivated CD-FBS were added to 2 × 10 ⁴ cells in a 48-well culture plate. / Well, and NF-κB activating receptor ligand: RANKL (manufactured by R & D Systems) is added to a final concentration of 100 ng / ml, so that the final concentration is 100 mg / ml. After adding the Momotake extract obtained in Example 2 dissolved in DMSO at a concentration of 0.1% (v / v), it was cultured at 37 ° C. under 5% carbon dioxide.

  As control groups, two groups were set: (1) RANKL added, no makomotake extract added, (2) RANKL added, no makomotake extract added.

  After changing the medium on the fourth day of culture, the medium was removed on the seventh day, and the cells were washed twice with PBS and then stained with a tartrate-resistant phosphatase staining kit (Sigma). Two groups were compared. Since osteoclasts are stained red with the above staining kit, the differentiation / proliferation state of osteoclasts can be observed by observing the staining state of the cells.

  FIG. 1 is a photograph showing the results of the control group 1 (no RANKL added, no makomotake extract added), FIG. 2 is a photograph showing the results of the control group 2 (with RANKL added, no makomotake extract added), and the test group (with RANKL added) FIG. 3 shows a photograph showing the results of the addition of Momotake extract.

  As a result, stained osteoclasts (the blackened portion in the figure) were clearly observed in the RANKL-added group and the No-Macomotake extract-added group in FIG. In contrast, no stained osteoclasts were observed in the RANKL-free group and the makomotake extract-free group shown in FIG. 1 and the RANKL-added and makomotake extract-added group shown in FIG.

  Therefore, a strong inhibitory activity against osteoclast differentiation / proliferation was observed by the addition of Mokomyotake extract.

According to a conventional method, hard capsules were filled with 230 mg / capsule according to a conventional method to produce hard capsule foods. The fish bone calcium used the brand name "baking Bonical" (made by Yaizu Suisan Chemical Co., Ltd.).

  A jelly beverage was produced according to a conventional method with the formulation shown in Table 2. This jelly beverage was very easy to drink without feeling the flavor of makomotake and excellent in texture.

  A milk beverage was produced according to a conventional method with the formulation shown in Table 3. This milk drink was very easy to drink without causing precipitation and without the taste of makomotake.

  The osteoclast differentiation / proliferation-inhibiting composition of the present invention can be used for the prevention / treatment improvement of osteoporosis by ingesting the composition as it is or in a food or drink product.

It is a photograph showing the positive state of differentiation / proliferation inhibitory activity of osteoclasts in a control group (RANKL-free addition, makomotake extract-free addition). It is a photograph showing the negative state of the osteoclast differentiation / proliferation inhibitory activity in a control group (RANKL added, no Macomotake extract added). It is a photograph showing the positive state of the differentiation / proliferation inhibitory activity of the osteoclast in the test group (RANKL addition, Makomotake extract addition).

Claims (4)

  A composition for inhibiting osteoclast differentiation / proliferation, comprising an extract of makomotake and / or pulverized mash of bamboo shoots as an active ingredient.   The composition for inhibiting osteoclast differentiation / proliferation according to claim 1, wherein the extract of makomotake is obtained by extracting makomotake with chloroform or ethanol.   A food or drink comprising the osteoclast differentiation / proliferation-inhibiting composition according to claim 1 or 2.   The food / beverage products of Claim 3 which contains the said osteoclast differentiation and proliferation inhibitory composition 0.5-70 mass% in conversion of dry makomotake.