JP2009096718A - Osteoclast differentiation inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a drug useful in preventing or improving bone resorptive diseases such as osteoporosis or a food useful in preventing or improving bone resorptive diseases such as osteoporosis. <P>SOLUTION: An osteoclast differentiation inhibitor and a bone resorption inhibitor have Myrica cerifera, Kaempferia galanga, Pimenta officinalis or an extract thereof as an active ingredient. An agent for preventing or improving bone resorptive diseases and the food for preventing or improving bone resorptive diseases have Myrica cerifera, Kaempferia galanga, Pimenta officinalis or an extract thereof as an active ingredient. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to an osteoclast differentiation inhibitor, a bone resorption inhibitor, or a pharmaceutical or food containing them for preventing or improving bone resorbable diseases such as osteoporosis.

  Bone tissue is maintained by a dynamic cycle of destruction (bone resorption) and formation (bone formation). Osteoclasts and osteoblasts are responsible for the bone resorption and bone formation, respectively. Osteoclasts involved in bone resorption are giant multinucleated cells, and are stimulated by soluble RANKL (soluble receptor activator of NF-κB ligand; sRANKL) and M-CSF (macropage colony stimulating factor) produced by osteoblasts. It is known to differentiate from mononuclear osteoclast precursor cells and then mature. Under normal conditions, the balance of activity is maintained between osteoclasts and osteoblasts (coupling), thus maintaining bone shape and quantity. If this balance tilts in the negative direction (bone resorption), bone mass decreases and eventually causes osteoporosis.

  Osteoporosis is a skeletal disease characterized by a decrease in bone strength associated with a decrease in bone mass. Osteoporosis is common in postmenopausal women and older men, about 30% in women in their 60s, more than about 40% in their 70s, about 10% in men in their 60s, It has been reported that about 20% of people in their 70s have osteoporosis. Osteoporosis that occurs with such aging is called evolutionary osteoporosis, which is postmenopausal osteoporosis that develops after menopause and senile osteoporosis that develops in old age and senile osteoporosis. Separated. Regressive osteoporosis is widely known as a disease that significantly lowers the QOL (quality of life) of affected individuals.

  Aggressive intake of calcium is important for the prevention and treatment of osteoporosis. In order to promote the intake of calcium, foods containing DFAIII (twintose), FOS (fructooligosaccharide), CPP (casein phosphopeptide), calcium citrate malate, etc. have been used (Patent Documents 1 to 4). . However, calcium intake from these foods alone is not sufficient to prevent bone loss.

  In order to effectively prevent bone loss, it is desirable to actively suppress bone resorption. As drugs that suppress bone resorption, bisphosphonate preparations, selective estrogen receptor modulators (SERM), calcitonin preparations, and the like have been generally used (Non-patent Documents 1 to 3). On the other hand, quercetin contained in onions and the like, curcumin contained in turmeric, and β-cryptoxanthin contained in Satsuma mandarin have been reported as food ingredients having the ability to suppress bone resorption (Non-Patent Documents 4 to 6).

  Shirokoyamamomo is a plant of the family Myricaceae and has been known to have a medicinal effect on toothache (Non-patent Document 7). Banturmeric is a perennial herb of Zingiberaceae Kaempferia, and is also called “Ban-turmeric” or “Sanna”. Allspice is an evergreen tree of Myrtaceae, also known as Hyakumikosho, Sanko, Pimento, Jamaican pepper, and immature fruits are dried and used as a spice . As other functions of allspice, an antioxidant action and an anti-aging action are known (Patent Documents 5 and 6).

However, the bone resorption inhibitory action and osteoporosis effect of these plants have not been reported so far.
Japanese Patent Laid-Open No. 11-43438 JP 7-252156 A JP 7-308172 A JP-A-1-156985 JP 2003-095915 A JP 2004-2237 A Kushida K. Et al. Bone Miner. Metab. (2004) 22, 462-468 Ohta H. et al. : Clin. Calcium (2004) 14, 73-80 Hosoi T. : CLINICAL CALCIUM (2004) 14 (11), 83-86 Wattel A.E. Et al. Cell. Biochem. (2004) 92, 285-295 Bharti AC. Et al. Immunol. (2004) 172, 5940-5947 Uchiyama S. et al. And Yamaguchi M. et al. : Biochem. Pharmacol. (2004) 67, 1297-1305 Keiji Uehara: Tree Illustration (1959), Ariake Shobo, Volume 1, 594

  The present invention relates to the provision of pharmaceuticals or foods useful for the prevention or improvement of bone resorbable diseases such as osteoporosis.

  As a result of studying the bone resorption inhibitory action and osteoporosis effect of various substances, the present inventors have found that white yam, van turmeric and allspice and their extracts have an osteoclast differentiation inhibitory action, and inhibit bone resorption. It was also found useful for the prevention or improvement of bone resorbable diseases such as osteoporosis.

  That is, the present invention relates to an osteoclast differentiation inhibitor containing, as an active ingredient, white bayberry, van turmeric, allspice or an extract thereof.

  Moreover, this invention relates to the bone resorption inhibitor which uses white bayberry, a turmeric, allspice, or those extracts as an active ingredient.

  The present invention also relates to a preventive or ameliorating agent for bone resorbable diseases comprising white bayberry, van turmeric or allspice or an extract thereof as an active ingredient.

  Furthermore, this invention relates to the food for prevention or improvement of the bone resorbable disease containing white bayberry, turmeric or allspice, or those extracts.

  ADVANTAGE OF THE INVENTION According to this invention, the pharmaceutical or foodstuff for preventing or improving bone resorbable diseases, such as osteoporosis, is provided.

As used herein, “white peach” refers to “ Myrica cerifera L ”.

As used herein, “van turmeric” means “ Kaempferia galanga L ” of the genus Kaempferia , and is also referred to as “Ban turmeric” or “Yanna” (Sanna). Bang turmeric is distinguished from turmeric belonging to the genus Curcuma. Curcumin, an active ingredient of bone resorption contained in Curcuma turmeric, is hardly contained in Van turmeric (Dictionary of Natural Products, Chapman & Hall, London, 1994).

  As used herein, “allspice” is an evergreen tree of the Myrtaceae family, also known as Hyakumikosho, Sanko, Pimento, and Jamaican pepper.

  In the present invention, any part of the plant can be used. For example, any one of the above-mentioned whole tree, whole grass, root, rhizome, stem, branch, stem, leaf, bark, sap, resin, flower, fruit, seed, etc., and any combination thereof, Multiple can be used. Among these, it is preferable to use bark for white bayberry, to use rhizome for turmeric, and to use fruit for allspice.

  The plant can be used as it is or after being dried. Alternatively, the plant may be chopped or pulverized, or may be pasted.

  The extract of the present invention is obtained by extracting the plant according to a known method at room temperature or under heating, using an appropriate solvent and using an instrument such as a Soxhlet extractor as necessary. Can do. The resulting extract can be concentrated, diluted, or dried and further powdered or pasted as required. The extract can also be purified by removing contaminants according to known methods. The extract can also be subjected to treatments such as deodorization and decolorization according to known methods, if necessary.

  Solvents used in preparing the extract of the present invention include polar solvents, nonpolar solvents, or combinations thereof. Specifically, for example, water; alcohols (for example, methanol, ethanol, propanol, butanol, 1,3-butylene glycol, propylene glycol); ketones (for example, acetone, methyl ethyl ketone); esters (for example, methyl acetate) Ethyl acetate); linear and cyclic ethers (eg, tetrahydrofuran, diethyl ether); polyethers (eg, polyethylene glycol); halogenated hydrocarbons (eg, dichloromethane, chloroform, carbon tetrachloride); hydrocarbons ( For example, hexane, cyclohexane, petroleum ether); aromatic hydrocarbons (for example, benzene, toluene); pyridines; supercritical carbon dioxide; and fats and oils, waxes, and other oils. Of these, water, alcohol or a mixture thereof is preferred. Water, ethyl alcohol or 1,3-butylene glycol or a mixture thereof is particularly preferable.

  In an exemplary extraction procedure, the extract can be obtained by adding 100 ml of solvent to 10 g of plant and shaking for several hours under heating or standing at room temperature for several days and then filtering. If necessary, the extract is concentrated by an evaporator and then dissolved in another solvent.

  As shown in the Examples below, white bayberry, turmeric and allspice and their extracts suppress the differentiation of RAW264.7 cells into osteoclasts. RAW264.7 is a cell line derived from mouse macrophages and is known to differentiate and mature into osteoclasts in the presence of soluble RANKL and M-CSF without depending on co-culture with osteoblasts. It has been used for various functional analyzes (Hsu H. et al .: Proc. Natl. Acad. Sci. USA (1999) 96, 3540-3545; Nirupama K. et al .: Proc. Natl. Acad. Sci. USA ( 2000) 97, 7829-7834; Hotokezaka H. et al .: J. Biol. Chem. (2002) 277, 47366-47372; Garcia V. et al .: J. Biol. Chem. (2005) 280, 13720-13727). Therefore, the white bayberry, van turmeric and allspice of the present invention and their extracts can act as osteoclast differentiation inhibitor or bone resorption inhibitor. The plant of the present invention and the extract thereof are also a research reagent for suppressing osteoclast differentiation, for suppressing bone resorption, or for preventing or ameliorating a bone resorbable disease such as osteoporosis, or a human or animal Can be used as a food, pharmaceutical or quasi-drug for, or for producing them. When used in foods, pharmaceuticals, quasi drugs, etc., the plants of the present invention and their extracts may be used alone or in combination of two or more.

  Here, the term “inhibition of bone resorption” refers to decreasing or stopping the bone resorption rate.

  The term “bone resorbable disease” means a disease or condition resulting from bone resorption exceeding bone formation. Examples of such diseases or conditions include bone loss or osteoporosis, and associated pains such as back pain and back pain, bone deformation, reduced bone strength, fractures, and the like.

  When the osteoclast differentiation inhibitor or bone resorption inhibitor of the present invention is used as a food, in addition to general foods, beauty foods, foods for the sick, and nutritional functional foods for the purpose of preventing or improving bone resorbable diseases Or it can be set as functional foods, such as food for specified health. The food can be solid, semi-solid or liquid. Examples of foods include breads, noodles, confectionery, jelly, dairy products, frozen foods, frozen foods, other processed foods, beverages, soups, seasonings, nutritional supplements, and the like, and their raw materials. . The food may be in tablet form, pill form, capsule form, liquid form, syrup form, powder form, granule form and the like.

  When used as food, the plant of the present invention or an extract thereof may be used alone or in combination with other food materials. The food of the present invention may contain a food additive as necessary. Examples of food additives include solvents, oils, softeners, emulsifiers, preservatives, stabilizers, antioxidants, colorants, UV absorbers, humectants, thickeners, brighteners, sweeteners, flavorings, etc. Can be mentioned.

  The amount of the plant of the present invention or the extract thereof in food is usually 0.0001% by mass to 50% by mass, preferably 0.0003% by mass of the total mass of the food in terms of dry matter of the extract. % To 10% by mass, and more preferably 0.0005% to 5% by mass.

  When the osteoclast differentiation inhibitor or bone resorption inhibitor of the present invention is used as a pharmaceutical, it can be administered in any dosage form. Administration forms include oral and parenteral administration. Examples of the dosage form of the preparation for oral administration include tablets, pills, capsules, powders, granules, syrups, elixirs, liquids and the like. Examples of parenteral administration include injection, infusion, transdermal, transmucosal, nasal, inhalation, suppository, and bolus.

  In such formulations, the plants of the invention or their extracts may be used alone or in combination with a pharmaceutically acceptable carrier. Such carriers include, for example, excipients, binders, disintegrants, lubricants, diluents, osmotic pressure adjusters, pH adjusters, emulsifiers, preservatives, stabilizers, antioxidants, colorants, ultraviolet light Examples include absorbents, humectants, thickeners, brighteners, activity enhancers, anti-inflammatory agents, bactericides, flavoring agents, and flavoring agents.

  The amount of the plant of the present invention or the extract thereof in the preparation is usually 0.01% by mass to 100% by mass of the total mass of the preparation in terms of the dry matter of the extract, preferably 0.1% It is mass%-70 mass%, More preferably, it is 0.5 mass%-50 mass%.

  The dosage of the formulation may vary according to the patient's condition, weight, sex, age, or other factors. The preferred daily dose per adult is 0.002 to 2000 mg / Kg / day, preferably 0.02 to 200 mg / Kg / day, in terms of dry matter of the extract of the present invention. More preferably, it is 0.2 to 20 mg / Kg body weight / day. The formulation can be administered according to any dosage regimen. For example, continuous administration can be administered three times daily, twice daily, once daily, once every two days, once every three days, once a week, or at any period and interval .

  The pharmaceutical or food of the present invention may be prepared in combination with at least one calcium intake or absorption enhancer or bone metabolism improving agent. Examples of calcium intake or absorption promoters include calcium salts such as calcium carbonate, DFAIII (twintose), FOS (fructooligosaccharide), CPP (casein phosphopeptide), vitamin D and the like. Examples of the bone metabolism improving agent include calcium salts such as calcium carbonate, vitamin D, vitamin K, soybean isoflavone, collagen, MBP (milk basic protein) and the like.

  The following examples illustrate the invention in more detail.

Preparation of Extract To 10 g of white root of white bayberry (US), 100 ml of 50% ethanol aqueous solution was added, allowed to stand at room temperature for 7 days, and then the solution was filtered to obtain an extract (78 ml, evaporation) Residue 2.9 w / v%). In the same manner, 87 ml (evaporation residue 1.1 w / v%) of an extract was obtained from 10 g of rhizome of Banturmeric (made in China). Further, in the same procedure, an extract (solid content concentration: 1.33 w / v%) was obtained from 10 g of Allspice (obtained from Tochimoto Tenkaido).

Example 1: Osteoclast differentiation inhibitory effect of the extract of the present invention 1) Screening RAW264.7 (Dainippon Pharmaceutical, Cat. No. 06-071) was suspended in α-MEM containing 10% fetal bovine serum, A 48-well plate was seeded at a concentration of 3.0 × 10 ³ / well and cultured overnight at 37 ° C. in the presence of 5% CO ₂ . The extract obtained by the above procedure was added to the wells, diluted stepwise with a medium without the extract, and then pre-cultured for a certain time (1-2 days). To this preculture, mouse sRANKL 50 ng / ml and mouse M-CSF 50 ng / ml were added to induce osteoclast differentiation. After culturing for 3 days, the tartrate-resistant acid phosphatase (TRAP) activity of each well was measured by the absorbance method. The osteoclast differentiation-inhibiting activity of each extract is the TRAP activity of a well to which only 50% ethanol aqueous solution having the same volume as the extract is added as a positive control (100% differentiation), and the TRAP activity of the well not induced to differentiate. As a negative control (0% differentiation), it was expressed as a percentage.

2) Results White bayberry, van turmeric and allspice extracts suppressed the differentiation of RAW264.7 cells into osteoclasts in a dose-dependent manner. The results are shown in FIG. The vertical axis represents the relative TRAP activity of differentiated osteoclasts compared to the control, and the horizontal axis represents the concentration of each extract (residual equivalent). In any extract, it was observed that TRAP activity decreased with increasing concentration.

Example 2 Bone resorption inhibiting beverage The following composition was mixed to produce a fruit juice beverage.

Example 3 Bone Resorption Suppressing Food The following composition (1 tablet = 1000 mg) was tableted to produce a chewable tablet food.

It shows osteoclast differentiation-inhibiting activity of white bayberry, van turmeric and allspice extracts.

Claims (6)

  An osteoclast differentiation inhibitor comprising, as an active ingredient, white bayberry, van turmeric or allspice or an extract thereof.   Bone resorption inhibitor comprising white bayberry, turmeric or allspice or an extract thereof as an active ingredient.   A preventive or ameliorating agent for bone resorbable disease comprising white bayberry, van turmeric or allspice or an extract thereof as an active ingredient.   The preventive or ameliorating agent according to claim 3, wherein the bone resorbable disease is osteoporosis, bone loss or fracture.   A food for preventing or ameliorating a bone resorbable disease containing white bayberry, turmeric or allspice or an extract thereof.   The food according to claim 5, wherein the bone resorbable disease is osteoporosis, bone loss or fracture.

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