JP2009073768A - Novel compounds and osteoclast differentiation / proliferation inhibitors - Google Patents

Abstract

A novel prophylactic / therapeutic agent for osteoporosis is provided.
A compound of the following formula:

[Effect] It can be used as a raw material for foods, drinks, pharmaceuticals and cosmetics.
[Selection figure] None

Description

  The present invention relates to, for example, a novel compound that can be used as a raw material for foods and drinks, pharmaceuticals, and cosmetics, and an osteoclast differentiation / proliferation inhibitor.

  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 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 below discloses a composition for promoting bone formation and preventing bone mineral content reduction, which contains 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, the present invention can be expected to have a sufficient effect of preventing or treating osteoporosis by inhibiting osteoclast differentiation / proliferation and suppressing bone resorption, and is used as a raw material for foods, beverages, pharmaceuticals, and cosmetics. It is an object of the present invention to provide a novel compound and an osteoclast differentiation / proliferation inhibitor that can be used.

  Makomotake is used as a raw material for traditional Chinese medicine, and various physiologically active effects such as suppression of dry mouth, fever reduction, bowel regulation, digestion and detoxification are known. The present inventors have investigated the further physiological activity of Momotake, which has been confirmed to be safe by abundant dietary experiences, and found the effect of preventing and treating osteoporosis as a new physiological activity of Momotake. As a result of further research on the physiologically active component, the present invention has been achieved.

  That is, the novel compound of the present invention is represented by the following chemical formula (1).

  It is preferable that the novel compound of this invention is used as a raw material of food-drinks, a pharmaceutical, or cosmetics.

  On the other hand, the osteoclast differentiation / proliferation inhibitor of the present invention contains a compound represented by the following chemical formula (1) as an active ingredient.

  In addition, the osteoclast differentiation / proliferation inhibitor of the present invention preferably contains a composition prepared from an organic solvent extract of Makomotake.

  Moreover, it is preferable that the osteoclast differentiation / proliferation inhibitor of the present invention contains a composition prepared from an organic solvent extract of the cervix of the fungus of Makomotake.

  The novel compound of the present invention can be used, for example, as a raw material for foods and drinks, pharmaceuticals, cosmetics and the like, and can be expected to have an effect of preventing or treating osteoporosis.

  Further, the osteoclast differentiation / proliferation inhibitor of the present invention contains a component derived from Makotake that has been confirmed to be safe by abundant dietary experience as an active ingredient, and is highly safe and further sufficient. Expected to be effective in preventing and treating osteoporosis.

  The novel compound of the present invention is represented by the following chemical formula (1).

  The compound represented by the above chemical formula (1) (hereinafter referred to as “compound (1)”) is a compound that is soluble in an organic solvent, and can be obtained, for example, by purifying a solvent extract of Makomotake.

  That is, first, the mushroom that is the raw material for extraction is dried or pulverized as necessary. Especially, since the content of the above-mentioned compound (1) is high in the cervix of Momotake, the recovery rate of the above-mentioned compound (1) can be improved by using the cervix of Momotake as an extraction raw material. Here, Makomotake is a cultivated stalk of Macomo, an aquatic plant belonging to the genus Macomo and Perennials, that grows hypertrophic by Aspergillus oryzae and is cultivated from East Asia to Southeast Asia, mainly in China and Taiwan. Vegetables. In recent years, cultivation has been carried out in Japan, and these commercial products can be easily obtained.

  Next, this extract raw material, Momotake, is immersed in an extraction solvent at room temperature to under heating to obtain a solvent extract. The extraction solvent is not particularly limited, and examples thereof include lower alcohols (methanol, ethanol, etc.), acetone, acetonitrile, chloroform, dichloromethane, ethyl acetate, and the like. These can be used alone or in combination of two or more. . Particularly preferred extraction solvents include methanol, ethanol, acetone, dichloromethane, and mixed solvents thereof.

  Since this extraction solvent often contains a large amount of contaminants, for example, makomotake is soaked in a water-soluble organic solvent such as lower alcohols or acetone at room temperature to under heating to After obtaining the solvent-extracted fraction, separate the fat-soluble component and the water-soluble component using a water-insoluble organic solvent such as ethyl acetate, hexane, chloroform, or dichloromethane, and collect the water-insoluble organic solvent-extracted fraction. It is preferable. By carrying out like this, the solvent extract (non-water-soluble organic-solvent extraction fraction) of Makotake which has very few impurities can be obtained. Moreover, this solvent extract may be concentrated as needed, and may be further dried.

  Next, the solvent extract is purified by column separation purification, recrystallization, or the like, whereby the compound (1) can be obtained.

  For the purification of the solvent extract, first, a crude product obtained by primary purification using chromatography is subjected to at least one method selected from recrystallization, column chromatography, and thin layer column chromatography. Subsequent purification is preferred. By carrying out like this, the recovery rate of the said compound (1) can be improved.

  The chromatography used for the primary purification is not particularly limited, and conventional general chromatography such as normal phase column chromatography, reverse phase column chromatography, size exclusion column chromatography, and ion exchange column chromatography may be used. it can. The primary purification method is not particularly limited. For example, when chromatography using silica gel as a carrier, a water-insoluble organic solvent such as chloroform, dichloromethane, benzene, ethyl acetate, hexane, and a mixed solvent thereof, Furthermore, it is preferable to add an appropriate amount of a water-soluble organic solvent such as methanol, ethanol, acetone, etc., and change the ratio to elute while increasing the polarity sequentially. Hexane / ethyl acetate, hexane / dichloromethane, dichloromethane / acetone, chloroform / acetone A method using a mixed solvent of chloroform / methanol is particularly preferable.

  Column chromatography used for the secondary purification includes normal phase column chromatography, diol column chromatography, reverse phase column chromatography, size exclusion column chromatography and the like, and purification conditions such as carrier and elution solvent are various chromatography. Can be selected as appropriate. These column chromatography can be employed singly or in combination, and can be appropriately applied to open columns, HPLC columns and the like.

  Examples of the thin layer chromatography used for the secondary purification include those using a reverse phase carrier or normal phase carrier or a chemically modified carrier modified with an amine or an optically active substance. It can be selected appropriately according to the chromatography.

  The novel compound of the present invention (compound (1)) is a component derived from Makotake that has been confirmed to be safe by abundant dietary experience, and its safety has already been proven, and is a raw material for various foods, pharmaceuticals, and cosmetics. Can be used as

  Examples of the food and drink include (a) soft drinks, carbonated drinks, fruit drinks, vegetable juices, lactic acid bacteria drinks, milk drinks, soy milk, mineral water, tea-based drinks, coffee drinks, sports drinks, alcoholic drinks, jelly drinks, and the like. (B) Processed vegetable products such as tomato puree, canned mushrooms, dried vegetables, pickles, (c) Processed fruit products such as dried fruits, jam, fruit puree, canned fruits, (d) curry powder, wasabi, Spices such as ginger, spice blend, seasoning powder, (e) noodles (including raw noodles, dried noodles) such as pasta, udon, soba, ramen, macaroni, (f) breads such as bread, confectionery bread, cooking bread, donut , (G) Alpha-ized rice, oatmeal, rice cake, batter flour, etc. (h) Baked confectionery, biscuits, rice confectionery, candy, chocolate, chewing gum , Snacks, frozen confectionery, candied confectionery, Japanese confectionery, Western confectionery, half confectionery, pudding, ice cream and other confectionery, (i) beans products such as red beans, tofu, natto, kinakome, yuba, boiled beans, peanuts, (j ) Honey, royal jelly processed foods, (k) meat products such as ham, sausage, bacon, (l) dairy products such as yogurt, pudding, condensed milk, cheese, fermented milk, butter, ice cream, (m) processed egg products, (N) Processed fish such as dried fish, sea bream, chikuwa, fish sausage, processed seaweed such as dried seaweed, kelp, and boiled fish, processed fish eggs such as octopus, scallop, salmon roe, and sea bream, (o) dashi stock, soy sauce, Condiments such as vinegar, mirin, consomme base, Chinese base, concentrated soup stock, dressing, mayonnaise, ketchup, miso, salad oil, sesame oil, linoleic oil, diacyl glycero , Edible fats and oils such as Benibana oil, (p) Cooking, semi-cooked foods such as soups (including powder and liquid), side dishes, retort foods, chilled foods, semi-cooked foods (eg cooked rice, crab Tamanomoto) and the like. And as addition amount to food / beverage products, if an effective amount can be taken, it will not specifically limit, What is necessary is just to mix | blend suitably so that it may not impair the taste and quality stability of food / beverage products, For example, 5-500 ppm mass. Preferably, it may be blended so that the mass becomes 10 to 100 ppm.

  Examples of the cosmetics include lotions, emulsions, lotions, gels, foundations, creams, facial cleansers, body washing agents, and the like. And as addition amount to cosmetics, 5-500 ppm mass is preferable, and 10-100 ppm mass is more preferable. If the addition amount is less than the above range, sufficient physiological activity effect cannot be expected, and if it is more than the above range, there may be a problem in quality stability.

  Examples of the pharmaceuticals include osteoclast differentiation / proliferation inhibitors, immunostimulators, melanin production inhibitors, and the like. Examples of the form include dosage forms such as liquids, powders, tablets, pills, fine granules, granules, capsules, jelly, chewable and paste.

  Next, the osteoclast differentiation / proliferation inhibitor of the present invention will be described.

  The osteoclast differentiation / proliferation inhibitor of the present invention contains a compound represented by the following chemical formula (1) (compound (1) as an active ingredient.

  And, the osteoclast differentiation / proliferation inhibitor of the present invention preferably contains a composition prepared from an organic solvent extract of Makomotake, a composition prepared from an organic solvent extract of the fungus colony of Makomotake It is more preferable to contain.

  The compound (1) is a component contained in Makotake and is a compound that is soluble in an organic solvent. For this reason, the organic solvent extract of Makomotake contains the compound (1), and a high osteoporosis prevention / treatment effect can be obtained. Especially, since the organic solvent extract of the fungus colony of Macomotake contains a large amount of the above compound (1), the composition prepared from the organic solvent extract of the fungus colony of Makomotake is used. Thus, the content of the active ingredient can be improved, and a more excellent osteoporosis prevention / treatment effect can be obtained.

  In addition, the extract obtained from the organic solvent extract of Makomotake usually contains about 1 ppm to 1000 ppm of the above compound (1) per solid content.

  In addition, the osteoclast differentiation / proliferation inhibitor of the present invention preferably further contains one or more selected from isoflavones, collagen peptides, lactoferrin, casein phosphopeptides, fish bone calcium and shellfish calcium. These components are known to have osteoporosis prevention / treatment effects, and a synergistic effect can be expected. Moreover, inorganic salts, organic acids, carbohydrates, proteins, peptides, amino acids, and lipids can be appropriately blended as necessary.

  Examples of the form of the osteoclast differentiation / proliferation inhibitor of the present invention include dosage forms such as solution, powder, tablet, pill, fine granule, granule, capsule, jelly, chewable and paste. The effective intake of the compound (1) is preferably 0.01 to 5 mg, more preferably 0.05 to 0.5 mg per kg body weight.

  Hereinafter, the novel compound and osteoclast differentiation / proliferation inhibitor of the present invention will be described in detail.

For identification of the compounds, ESI / TOFMS spectrum analysis, infrared absorption (IR) spectrum analysis, nuclear magnetic resonance (NMR) spectral analysis, DEPT spectrum ^analysis, 1 ^H- 1 H COZY spectrum analysis, HMQC spectrum analysis, HMBC spectrum The analysis was performed.

  ESI / TOFMS spectral analysis was performed in positive ion mode.

  Infrared absorption (IR) spectrum analysis was performed by the KBr method.

Nuclear magnetic resonance (NMR) spectrum analysis was performed under the conditions of 500 MHz for ¹ H-NMR, 125 MHz for ¹³ C-NMR, and deuterated chloroform.

<Example 1>
[New compound]
Makomotake was cut into leaves and fungal parts, and the fungal parts (30.6 kg) were pulverized with a pulverizer and extracted sequentially with 85% EtOH and acetone. Thus was obtained extract partitioned solvent fraction with CHCl ₃ and EtOAc and water to give CHCl ₃ soluble part, EtOAc soluble portion, a water-soluble portion. When a part of the obtained fraction was subjected to an osteoclast formation inhibition test, activity was confirmed in the CHCl ₃ soluble part and the EtOAc soluble part. Therefore, 7.2 g of the EtOAc soluble part was subjected to flash column chromatography (silica gel 60 N (350 g), φ4 cm × 60 cm), and CH ₂ Cl ₂ : acetone = 10: 0, 9: 1, 6: 4, Elution was performed stepwise with a solvent of 2: 8, 0:10, 100% MeOH to obtain elution fractions MAK-E1 to MAK-E26. When these were subjected to the osteoclast formation inhibition test described later, the activity was confirmed in MAK-E3,4,9,11,13,24. Of these, MAK-E4 (135.8 mg) was subjected to flash column chromatography (silica gel 60 N, (200 g), φ2.5 cm × 60 cm), hexane: CH ₂ Cl ₂ = 6: 4,100% CH ₂ Elution was performed stepwise with a solvent of Cl ₂ , CH ₂ Cl ₂ : MeOH = 1: 1, 100% MeOH to obtain elution fractions MAK-E4-1 to MAK-E4-7. The MAK-E4-6 (16.3 mg) was subjected to preparative TLC (silica gel 60 F ₂₅₄ , 20 × 20 cm) and developed with hexane: CH ₂ Cl ₂ = 6: 4, and the developed spot fraction MAK-E4 -6-1 to MAK-E4-6-10 were obtained. In addition, MAK-E-4-7 (76.7 mg) was subjected to preparative TLC (silica gel 60 F ₂₅₄ , 20 x 20 cm) and developed with hexane: CH ₂ Cl ₂ = 6: 4, MAK-E4-7-1 to MAK-E4-7-10 were obtained. When each fraction was compared by TLC and NMR, MAK-E4-6-3,4 and MAK-E4-7-3,4,5 were similar fractions. -1 '. Similarly, MAK-E4-6-5,6 and MAK-E4-7-6,7 are combined into MAK-E4-7-2 ', MAK-E4-6-7,8,9, and MAK- E4-7-8 and 9 were combined into MAK-E4-7-3 '. MAK-E4-7-1 ′ (10.2 mg) was subjected to Sep pak (silica gel), and the obtained elution part was subjected to HPLC (Senshu pak AQUASIL SS-5251) using a normal phase column, hexane: CHCl3 = Elution was performed at 6: 4 to obtain elution fractions MAK-E4-7-1′-1 to MAK-E4-7-1′-13. The eluted fraction MAK-E4-7-1'-2 (5.3 mg) was subjected to sep pak (silica gel) and eluted with 100% hexane to obtain a non-adsorbed part and an adsorbed part. However, since the obtained fraction was small in quantity and difficult to analyze for chemical structure identification, it was extracted again from the cervix of the genus Momotake.

  That is, the osteoclast formation inhibitory active substance was isolated as described below using the MAK-E4-7-1'-2 fraction as an index.

The fungus cervix (39.0 kg) was pulverized with a pulverizer and extracted successively with 85% EtOH and acetone. A hexane soluble part (40.0 g) was extracted from the obtained extract and subjected to flash column chromatography (silica gel 60 N, (350 g), φ4 cm × 60 cm) in the same manner as above, and the eluted fraction MAK- H1 to MAK-H26 were obtained. When each fraction was compared by TLC and NMR using the above MAK-E4-7-1′-2 as an index, MAK-H6 to MAK-H10 were similar fractions, and therefore MAK-H6 (107.0 mg). This was subjected to preparative TLC (silica gel 60 F ₂₅₄ , 20 × 20 cm), developed with hexane: CH ₂ Cl ₂ = 6: 4, and developed spot fractions MAK-H6-1 to MAK-H6-12 were obtained. Obtained. When each fraction was compared by TLC and NMR, MAK-H6-6,7,8,9,10 were similar fractions, so they were collectively referred to as MAK-H6-6-2 '. MAK-H6-6-2 '(25.9 mg) was subjected to Sep pak (silica gel), and the resulting elution part was subjected to HPLC (Senshu pak AQUASIL SS-5251) using a normal phase column and eluted with 100% hexane. Elution fractions MAK-H6-6-2′-1 to MAK-H6-6-2′-40 were obtained. Of these, MAK-H6-6-2'-27,28,29,30,31,32,33 were combined into MAK-H6-6-2'-27 'using TLC and NMR as indicators, and MAK-H6- 6-2'-34,35,36,37,38,39,40 was combined into MAK-H6-6-2'-34 '. MAK-H6-6-2'-27 'was subjected to HPLC under the same conditions as above to obtain elution fractions MAK-H6-6-27'-1 to MAK-H6-6-27'-20. Since the amount decreased at this stage, MAK-H6 (106.0 mg) was again subjected to preparative TLC under the same conditions as shown above, and five developed spot fractions MAK-H6-1 (2) to MAK- H6-5 (2) was obtained. Of these, MAK-H6-4 (2) (58.0 mg) was subjected to Sep pak (silica gel) and normal phase column HPLC (Senshu pak AQUASIL SS-5251) under the same conditions as above, and the eluted fraction MAK-H6- 4 (2) -1 to MAK-H6-4 (2) -14 were obtained. MAK-H6-6-2'-34 ”(24.0 mg) of the elution fraction MAK-H6-4 (2) -10,11 and the above MAK-H6-6-2'-34 ' It was subjected to a hydrophobic filter and eluted with 2-propanol, from which a novel compound (compound (1)) (2.1 mg) represented by the following formula (1) was isolated from the 2-propanol insoluble part.

  Below, it shows about the result of identification of the said compound (1).

As shown in FIG. 1, compound (1) showed a molecular ion peak of [M + Na] +, at m / z 687 in ESI / TOFMS, and thus has a molecular weight of 664 and a molecular formula of C ₄₆ H ₈₀ O _2. It became clear. Further, as shown in FIG. 8, since absorption was observed at 1740 cm ⁻¹ in the IR spectrum, the presence of a carboxyl group was clarified. ^{The 1} H-NMR spectrum showed a characteristic peak for steroids (Fig. 3), but the number of carbon atoms determined by ESI / TOFMS was 46, suggesting that this compound is a fatty acid ester of steroids. It was done. In the ¹³ C-NMR spectrum shown in FIG. 4, a peak of δ173.5 derived from a carboxyl group was observed. The presence of five quaternary carbons was suggested from the DEPT spectrum shown in FIG. From the correlation of the HMQC spectrum shown in FIG. 6, the assignment of C and the proton bonded thereto could be clarified. From the COZY spectrum shown in FIG. 7, correlations were confirmed between H-20 and H-21, and H-22 and H-23. From the HMBC spectrum shown in FIG. 8, H-3 and C-1 ′, H-3 ′ and C-2 ′, 3 ′, H-18 ′ and C-16 ′, 17 ′, H-1 and C-3 , 5, H-2 and C-3, H-4 and C-3, 5, H-6 and C-8, H-7 and C-9, 14, H-11 and C-9, H-12 And C-9,13, H-14 and C-12, H-15 and C-14, H-16 and C-13, H-17 and C-13, H-18 and C-12,13,14 , 17, H-19 and C-1,5,9,10, H-20 and C-17, H-21 and C-17,20,22, H-22 and C-17, H-26 and C Correlation between -23,24,25, H-27,28 and C-24,25 was confirmed, and a partial structure corresponding to the side chain of the steroid was confirmed. In addition, the correlation between H-3 and C-1 ′, H-3 ′ and C-2 ′, 3 ′, H-18 ′ and C-16 ′, 17 ′, which are shifted to a lower magnetic field than the hydroxyl group, was confirmed. A partial structure corresponding to an ester-linked fatty acid was confirmed.

  Based on the above, it was determined that the novel compound obtained from the cervix of the mushroom was Episterol stearate (steroid stearate) represented by the above formula (1).

<Test Example 1>
[Evaluation of osteoclast differentiation / proliferation inhibitory activity]
The osteoclast differentiation / proliferation inhibitory activity of the compound (1) was evaluated. The osteoclast formation inhibitory activity test used a co-culture method of mouse-derived bone marrow cells and osteoblast-like stromal cells. Osteoclast formation inhibitory activity was evaluated by counting the number of osteoclasts (TRAP positive multinucleated cells) formed.

・ Cultivated culture Bone marrow cells collected from tibia and femur of mice (female 5-7 weeks old) 1.3 × 10 ⁸ cells and osteoblast-like stromal cells 1 × 10 ⁶ cells collected from the skull and cultured The suspension was suspended in 7.2 ml of (10% FBS-containing α-MEM), and dispensed into a 48-well plate at 150 μl / well.

As a sample solution of the above compound (1), the solution obtained in Example 1 was dissolved in DMSO and hexane so as to be 100 mg / ml, and further diluted with a culture solution to prepare a dilution series. 50 μl of sample solution of each concentration was added to the culture solution together with 50 μl of 1,25 (OH) ₂ VD ₃ (20 ng / ml) to make a total volume of 250 μl / well. Cultivate in a CO ₂ incubator (37 ° C, CO ₂ concentration 5.0%). On the third day of culture, remove 100 μl of the supernatant from each well to add the sample. The sample solution and 50 μl each of 1,25 (OH) ₂ D ₃ (40 ng / ml) were added and cultured again.

・ Count of TRAP-positive multinucleated cells
After culturing for about 1 week, the culture solution was removed, washed with PBS, and then added with 10% formalin-containing PBS solution at 0.5 ml / well and fixed for 10 minutes. Next, EtOH was added at 0.5 ml / well and re-fixed for 1 minute. Thereafter, it was dried, 0.3 ml / well of TRAP reaction solution was added, and staining was performed at room temperature for 30 minutes. After staining, it was washed with distilled water (0.5 ml / well) and dried. Cells that were TRAP-positive and had 2 or more nuclei were counted as TRAP-positive multinucleated cells, and the number per well was counted. The TRAP reaction solution was prepared by dissolving 1.5 mg of Naphthol as-mx phosphate in 150 ml of N, N-dimethylformaldehyde, and adding 15 mg of fast red violet lb salt to 15 ml of buffer (0.1% containing 50 mM Na tartrate). The solution added to M Na acetate buffer (pH 5.0) was mixed and prepared.

・ MTT assay
The number of viable cells after the co-culture was confirmed by MTT assay. Specifically, without removing the culture solution after co-culture, 1 mg / ml MTT reagent was added at 125 μl / well. Next, after culturing at 37 ° C. under a CO ₂ concentration of 5.0% for 2 hours using a CO ₂ incubator, the culture solution was removed, 100 μl / well of dimethyl sulfoxide was added, and the absorbance (λ = 570 nm) was measured. The MTT reagent was prepared by dissolving 3- (4,5-dimethyl-2-thiazoly) -2,5-diphenyl-2H-tertazolium bromide in water to 1 mg / ml.

  The results are summarized in FIG. As apparent from FIG. 9, the compound (1) did not affect the cell viability at a concentration of 50 μg / ml, and significantly suppressed the formation of osteoclasts.

<Production example 1>
According to a conventional method, the hard capsules were filled with 230 mg / capsule according to a conventional method to produce a hard capsule food. The fish bone calcium used the brand name "baking Bonical" (made by Yaizu Suisan Chemical Co., Ltd.).

<Production example 2>
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.

<Production Example 3>
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 novel compound of the present invention can be used as a raw material for foods and drinks, pharmaceuticals, cosmetics and the like.

  In addition, the osteoclast differentiation / proliferation inhibitor of the present invention can be used for the prevention / treatment improvement of osteoporosis by ingesting it as it is or in combination with foods, beverages, pharmaceuticals, cosmetics and the like.

It is an ESI / TOFMS spectrum of compound (1). It is the same IR spectrum diagram. It is the same ¹ H-NMR spectrum figure. It is the ¹³ C-NMR spectrum figure. It is a corresponding figure of the DEPT spectrum (upper part) and a ¹³ C-NMR spectrum (lower part). It is the same HMQC spectrum figure. It is the same ¹ H- ¹ H COSY spectrum figure. It is the same HMBC spectrum figure. 4 is a chart showing osteoclast differentiation / proliferation inhibitory activity of the compound (1).

Claims (5)

A novel compound represented by the following chemical formula (1).
The novel compound according to claim 1, which is used as a raw material for foods, drinks, pharmaceuticals or cosmetics. An osteoclast differentiation / proliferation inhibitor comprising a compound represented by the following chemical formula (1) as an active ingredient.
4. The osteoclast differentiation / proliferation inhibitor according to claim 3, comprising a composition prepared from an organic solvent extract of Momotake. 5. The osteoclast differentiation / proliferation inhibitor according to claim 4, comprising a composition prepared from an organic solvent extract of the fungus cervix of Macomotake.