JP2009227616A - Osteogenesis promoter containing oleuropein and/or hydroxytyrosol as active ingredient - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an osteogenesis accelerator which acts to promote osteogenesis and inhibit bone resorption. <P>SOLUTION: The osteogenesis promoter includes oleuropein and/or hydroxytyrosol as an active ingredient and a pharmaceutical composition includes this osteogenesis promoter as an active ingredient. Further, the oleuropein and/or hydroxytyrosol is used for producing the osteogenesis promoter. Furthermore, a food composition includes oleuropein and designates that it is to be used for improving osteodystrophy and bone metabolic disorders. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to an osteogenesis promoter. More specifically, the present invention relates to an osteogenesis promoter containing oleuropein as an active ingredient, a pharmaceutical composition containing the same, and a food composition.

  Currently, about 1/3 of the population over 65 years old suffers from osteoporosis in Japan. The fact that about 80% of patients are postmenopausal women is thought to be due to changes in the amount of sex hormones after menopause and the fact that bone mass is smaller than men.

  The osteoporosis is a disease in which the bone mass per volume, that is, the bone density decreases, and the risk of fracture increases, and the frequency of onset increases with age. Osteoporosis has a very high risk of fractures, causing spinal compression fractures and femoral neck fractures.

  It is known that the cause of osteoporosis is composed of a plurality of genetic factors and factors related to lifestyle habits, and is caused by the interaction thereof. The treatment is mainly done by medication. If the increase in bone mass can be controlled by medication or other methods, it is possible to provide important knowledge for the treatment of diseases related to bone dysplasia represented by osteoporosis.

  Diseases related to abnormal bone metabolism such as osteoporosis are caused by a loss of the balance between bone resorption and bone formation. In normal bone tissue, bone formation and destruction are maintained in a well-balanced manner, and so-called “bone remodeling” is actively performed. In osteoporosis, since bone resorption exceeds bone formation, bone density decreases and bone vulnerability increases. Most of the osteoporosis that develops after menopause is considered to be caused by estrogen deficiency, and it is known that bone resorption is enhanced by lack of estrogen.

  Bone tissue contains cells called osteoblasts and osteoclasts that are involved in bone metabolism and function in “bone remodeling”. Osteoblasts are cells that play a central role in the bone formation process. Developmentally, it is derived from mesenchymal stem cells and differentiates into osteoblasts via preosteoblasts. Mature osteoblasts are involved in calcification. It is known that 10-20% of osteoblasts become bone cells.

  On the other hand, osteoclasts are cells that play a central role in the process of lysing the bone matrix. Developmentally, it is derived from hematopoietic stem cells and differentiates into mononuclear osteoclasts via precursor osteoclasts. Ultimately, mononuclear osteoclasts fuse to differentiate into multinucleated mature osteoclasts and participate in bone resorption.

  The chemical structures of oleuropein, hydroxytyrosol and tyrosol are shown in FIG. 1, respectively. These are the main active ingredients contained in olive extracts and are both flavonoids. Hydroxytyrosol is a degradation product of oleuropein, and tyrosol has a structure in which one OH group of hydroxytyrosol is removed.

  Hereinafter, “oleuropein” and “hydroxytyrosol” related to the present invention will be described.

  "Oleuropein" is a component contained in olives and a kind of polyphenol. With regard to oleuropein, its antiviral activity, antiprotozoal activity, and antibacterial activity have attracted attention. Non-patent document 1 and non-patent document 2 disclose a study using a bone resorption promoting model rat, and it is reported that reduction of bone density is improved by oral administration of oleuropein. However, these reports describe that components derived from oleuropein and olives suppressed the decrease in bone density by anti-inflammatory action. That is, the effect of oleuropein is only indicative of an anti-inflammatory effect, and no mention is made of bone formation and bone resorption (“bone remodeling”).

  “Hydroxytyrosol (3,4-dihydroxyphenylethanol)” is a degradation product of oleuropein, and is a kind of polyphenol contained in olive as well as oleuropein. Hydroxytyrosol exhibits an inhibitory activity on platelet aggregation, and is known to have a melanin production inhibitory action and a lipid peroxide production inhibitory action. Patent Document 1 discloses an external skin solvent and a bath preparation characterized by containing hydroxytyrosol as an active ingredient of a melanin production inhibitor or as an active ingredient of a lipid peroxide production inhibitor. However, the fact that hydroxytyrosol has an effect of promoting bone formation and suppressing bone resorption is the content disclosed for the first time in this case.

“Olive oil and its main phenolic micronutrient (oleuropein) prevent inflammation-induced bone loss in the ovariectomised rat.” British Journal of Nutrition, 2004, Vol. 92, p. 119-127 “Dose-response study of effect of oleuropein, an olive oil polyphenol, in an ovariectomy / inflammation experimental model of bone loss in the rat.” Clinical Nutrition, 2006, Vol. 25, p. 859-868 JP 2004-26836 A

  As described above, improving normal bone formation and promoting normal bone formation is a very important issue for the treatment of osteoporosis and further treatment for abnormal bone metabolism.

  Therefore, the main object of the present invention is to provide an osteogenesis promoter having an action of promoting osteogenesis.

  As a result of intensive studies for solving the above problems, the present inventors have found that oleuropein and / or hydroxytyrosol can suppress bone resorption. In addition, the inventors found that oleuropein and / or hydroxytyrosol promoted osteoblast activation and suppressed osteoclast formation, thereby promoting bone formation as a whole, and completed the present invention. I came to let you. That is, the present invention provides an osteogenesis promoter containing oleuropein and / or hydroxytyrosol as an active ingredient, and a pharmaceutical composition containing this osteogenesis promoter as an active ingredient. In addition, the present invention provides the use of oleuropein and / or hydroxytyrosol for producing an osteogenesis promoter. In addition, the present invention provides a food composition containing oleuropein and / or droxytyrosol and displaying that it is used to improve abnormal bone formation and abnormal bone metabolism. In the present invention, oleuropein and hydroxytyrosol can be derived from an olive extract.

  According to the present invention, an osteogenesis promoter having an action of promoting osteogenesis is provided. Since the active ingredient of this osteogenesis promoter can be derived from the olive extract, there is a high possibility that it has few side effects and is excellent in safety.

  As described later in detail in Examples 1 to 3, the present inventors have found that as an action of oleuropein and hydroxytyrosol, it promotes osteoblast differentiation activity or suppresses osteoclast formation. As a whole, it has been clarified that it has an action of promoting bone formation.

  Accordingly, the present invention provides an osteogenesis promoter containing this oleuropein and / or hydroxytyrosol as an active ingredient.

  The most notable point of the present invention is that, as will be described later in detail in Examples 2 and 3, the bone forming action by oleuropein and hydroxytyrosol is due to the activation of osteoblasts and the suppression of osteoclast formation. It was possible to clarify.

  Here, “activation of osteoblasts” in the present invention is used as a general term for the three stages of osteoblast proliferation, differentiation, and calcification. More specifically, “activation of osteoblasts” refers to a process in which osteoblasts proliferate and differentiate and mature to have calcification ability and promote bone formation. For osteoblast differentiation, alkaline phosphatase activity, which is a differentiation marker, can be used as an index, and calcification, which is the final stage of bone formation, can be measured by the amount of calcium deposited in mature osteoblasts. is there.

  In the present invention, “formation of osteoclasts” means that osteoclasts differentiate and become multinucleated mature osteoclasts. These can be evaluated by TRAP staining.

  The osteogenesis promoter according to the present invention is considered to promote osteogenesis by promoting osteoblast activity and suppressing osteoclast formation.

  More specifically, the inventors have demonstrated that at least oleuropein promotes osteoblast differentiation, and that oleuropein and hydroxytyrosol promote calcification by mature osteoblasts. On the other hand, it was proved that oleuropein and hydroxytyrosol inhibit the formation of osteoclasts.

  That is, according to the present invention, the action and effect of oleuropein and hydroxytyrosol could be proved at the cellular level. The specific targets of oleuropein and hydroxytyrosol are osteoblasts and osteoclasts, which provide very useful knowledge in the development of osteogenesis promoters.

  Therefore, the osteogenesis promoter according to the present invention can be applied to osteoporosis patients caused by increased bone resorption as compared with bone formation with respect to so-called “bone remodeling”, and can promote normal bone formation. Thus, it can be used to treat osteoporosis patients. It can also be used to prevent osteoporosis when applied to healthy individuals.

  In the osteogenesis promoter according to the present invention, oleuropein and hydroxytyrosol can be derived from a natural extract and a purified product of this extract. It is naturally possible to use chemically synthesized oleuropein and the like.

  When obtaining oleuropein and the like from natural product extracts, extraction from olives is particularly preferred. Olive is an evergreen tree native to the Mediterranean Sea, and olive oil obtained from its fruits has been edible for a long time. Regarding olive oil, it is widely known that oleic acid, which is a component thereof, has an action of reducing the amount of LDL cholesterol.

  It is already known that oleuropein and the like are contained in a large amount in olive leaves, and an extract containing oleuropein and the like can be obtained from olive leaves with water, an organic solvent such as alcohol or oil. As the organic solvent for olive extraction, methanol, ethanol, propanol, butanol, propylene glycol, 1,3-butylene glycol, glycerin, acetone, methyl ethyl ketone, ethyl acetate, ethers, chloroform, dichloromethane and the like can be used. . Furthermore, if the obtained extract is concentrated by a known method, oleuropein and the like can be obtained as a purified product.

  For olives, any part of its fruit, leaves, flowers, stems, and roots can be used, and these may be used in a raw state or in a dried product. Among these, oleuropein and the like are particularly abundant in olive leaves. Therefore, oleuropein and the like can be extracted by effectively using pruned and cut olive leaves. Further, when olive oil is squeezed from olive fruit, a large amount of plant water, treated water and pomace are generated, from which oleuropein and the like can also be extracted. Furthermore, since oleuropein and the like are also contained in olive oil, it may be extracted from olive oil, and in some cases, olive oil itself may be used.

  As mentioned above, olive oil has been edible for a long time, which indicates the high safety of the olive extract. Therefore, the osteogenesis promoter according to the present invention having oleuropein derived from olive extract as an active ingredient, and the pharmaceutical composition and food composition containing the same have the following advantages.

  The pharmaceutical composition according to the present invention includes, for example, tablets, capsules, elixirs, microcapsules, etc., which are sugar-coated as needed, or water or other pharmaceutically acceptable liquids. It can be used parenterally in the form of an injection such as a sterile solution or suspension. The pharmaceutical composition is required for the generally accepted formulation implementation of the osteogenesis promoter according to the present invention together with physiologically recognized carriers, flavoring agents, excipients, vehicles, preservatives, stabilizers, binders and the like. Manufactured in a unit dosage form. Additives that can be mixed into tablets, capsules and the like include binders such as gelatin, corn starch, tragacanth, gum arabic, excipients such as crystalline cellulose, corn starch, gelatin, alginic acid and the like. Leavening agents, lubricants such as magnesium stearate, sweeteners such as sucrose, lactose or saccharin, flavorings such as peppermint, red oil and cherry.

  The dosage of the pharmaceutical composition is determined in consideration of the type of tablet, the administration method, the age and weight of the administration subject, symptoms, and the like.

  Further, the food composition according to the present invention includes the above bone formation promoter, glucose, fructose, sucrose, maltose, sorbitol, stevioside, rubusoside, corn syrup, lactose, citric acid, tartaric acid, malic acid, succinic acid, lactic acid , L-ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerin, propylene glycol, glycerin fatty acid ester, polyglycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, gum arabic, carrageenan, casein , Gelatin, pectin, agar, vitamin B, nicotinamide, calcium pantothenate, amino acids, calcium salts, pigments, fragrances, preservatives, etc. It can be produced by. Further, it may be mixed with rice, wheat, corn, potato, sweet potato, soybean, kelp, wakame, tengusa and the like which are generally used as food materials.

  Examples of food compositions include health foods, functional foods, foods for specified health use, nutritional supplements, and light intestine nutritional foods.

  The food composition according to the present invention displays that it is used for promoting bone formation as necessary.

<Study on the effect of administration of oleuropein and its derivatives on bone density>
In order to examine the effect of administration of oleuropein on bone density, an experiment using a bone resorption model mouse by ovariectomy was performed.

  Balb / c mice (female, 6 weeks old) were used as animals. As a model for bone resorption, ovariectomized mice were used (“Ovariectomized section”). In addition, as a control experiment, a mouse subjected to a false operation (hereinafter referred to as “sham operation”) in which only the incision was performed without removing the ovary (“control group”) was used.

  Orophane, hydroxytyrosol, and tyrosol were orally administered as test substances every 10 days to mice in the ovariectomized and control groups so that the body weight was 10 mg / kg body weight, and BMD ( Bone Mineral Density) was measured. BMD was measured using X-ray CT for laboratory animals (LaTheta LCT-100, Aloka), and the bone density was calculated as the bone density of cancellous bone in the femur.

  The results are shown in “FIG. 2”. (A) When sham operation is performed (“control group”), (B) When the ovary is removed (“ovary removal group”), (C) The ovary is removed, and oleuropein, hydroxytyrosol, and tyrosol When administered, bone density is shown. The vertical axis represents the bone density value of cancellous bone at the inner portion 1.8 mm from the growth plate.

(A) The average value of bone density in mice subjected to sham operation (“control group”) was about 175 mg / cm ³ . (B) The average value of bone density in the mice from which the ovaries were only removed (“ovarian removal section”) was about 50 mg / cm ³ . In the ovariectomized group, the bone density was reduced by about 75% compared to the control group. On the other hand, (C) The average value of bone density in the mice from which ovaries were removed and oleuropein was administered was about 70 mg / cm ³ . The average bone density of the ovariectomized mice administered with hydroxytyrosol was about 90 mg / cm ³ . Bone density reduction was suppressed in ovariectomized mice dosed with oleuropein and hydroxytyrosol. In vivo, it was found that hydroxytyrosol is more effective in suppressing the decrease in bone density than oleuropein.

  From the above results, it was confirmed that oleuropein suppresses the decrease in bone density and has an effect on the recovery of bone density in the ovariectomized mouse. On the other hand, it has been proved for the first time that hydroxytyrosol has a more significant effect than oleuropein in suppressing the decrease in bone density.

<Examination of the effect of oleuropein and hydroxytyrosol on osteoblasts>
It is known that osteoblasts play an important role in bone formation by proliferation, differentiation, mineralization, and the like. Therefore, in order to know the mechanism of action of oleuropein and hydroxytyrosol on osteoblasts, in vitro experiments using osteoblast cell lines were performed.

  MC3T3-E1 cells, which are osteoblast cell lines, were used for experiments on osteoblast proliferation, differentiation, and calcification. As a control experiment, apigenin contained in the olive extract was used.

  Apigenin is the main active ingredient in olive extracts and is a type of flavonoid along with oleuropein. The chemical structure of apigenin is shown in (II) below.

  The culture conditions for the osteoblast cell line are shown below. Penicillin and streptomycin were added to α-MEM containing 10% fetal calf-derived serum at 50 units / ml and 50 μg / ml, respectively. Cells were cultured at 37 degrees in the presence of 5% carbon dioxide. When the osteoblast cell line became 70% confluent, the cells were peeled off using a 0.05% trypsin solution and plated on 96-well plates and 12-well plates for the following experiments.

(1) An MTT assay was performed to measure the proliferation rate of osteoblasts. Osteoblasts were seeded on a 96-well plate at 1 × 10 ³ cells / well, and oleuropein and hydroxytyrosol were added to each well to a concentration of 10 ⁻⁶ M to 10 ⁻⁴ M. After adding oleuropein and hydroxytyrosol, after 45 and 69 hours, add appropriate amount of 3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2H tetrazolium bromide as a substrate. Absorbance at 570 nm was measured using a photometer. As a control experiment, an experiment using apigenin, one of the same flavonoids, was conducted.

The results are shown in “FIGS. 3A, 3B, and 3C”. The vertical axis shows the absorbance at 570 nm. A larger value indicates higher proliferation activity. At 45 hours after the addition of oleuropein and hydroxytyrosol, there was no significant change in osteoblast proliferation. Even after 69 hours from the addition of oleuropein and hydroxytyrosol, there was no effect on osteoblast proliferation in the range of 10 ⁻⁶ M to 10 ⁻⁵ M. On the other hand, apigenin was found to suppress osteoblast proliferation activity even at 5 × 10 ⁻⁶ M.

From the above results, it was found that oleuropein and hydroxytyrosol had no significant effect on osteoblast proliferation at concentrations up to 10 ⁻⁵ M. Osteoblasts were not promoted by oleuropein and hydroxytyrosol.

(2) Alkaline phosphatase activity was measured in order to know the effect of oleuropein and the like on osteoblast differentiation. Osteoblasts were seeded on a 12-well plate at 3.8 × 10 ⁴ cells / well and cultured for 3 days. Thereafter, oleuropein was added to each well to a concentration of 10 ⁻⁶ M to 10 ⁻⁴ M. The medium containing oleuropein was changed every 3 days, and alkaline phosphatase activity was measured by the following method from the 12th to the 15th. As a control, an experiment using the aforementioned apigenin was performed.

  The cells were washed with 10 mM Tris-HCl (pH 7.2) solution, 1 ml of a solution containing 50 mM Tris-HCl (pH 7.2) and 0.1% Triton X-100 was added to each well, and sonicated for 15 seconds. . Alkaline phosphatase activity was performed according to a conventional method using p-nitrophenyl phosphate as a substrate. BCA protein assay kit (Pierce Chemical Co.) was used for protein measurement.

  The results are shown in “FIGS. 4A, 4B, and 4C”. The vertical axis shows alkaline phosphatase activity. It was found that the osteoblasts 14 days after adding oleuropein had higher alkaline phosphatase activity than the control. It was shown that alkaline phosphatase activity increased in a concentration-dependent manner with oleuropein. Further, no change in alkaline phosphatase activity was observed in osteoblasts supplemented with hydroxytyrosol. On the other hand, it was found that the alkaline phosphatase activity decreased in a concentration-dependent manner in osteoblasts added with apigenin.

  From the above results, it was shown that oleuropein promotes osteoblast differentiation in osteoblasts because it promotes alkaline phosphatase activity in a concentration-dependent manner. It was found that hydroxytyrosol is not significantly involved in osteoblast differentiation. Apigenin did not act to promote osteoblast differentiation. This suggests that the action of oleuropein is not a general effect on olive-derived flavonoids, but an effect specific to oleuropein.

(3) In order to know the effect of oleuropein and the like on the calcification of osteoblasts, the amount of calcium deposition was measured. Osteoblasts were seeded on a 12-well plate at 3.8 × 10 ⁴ cells / well and cultured for 3 days. Thereafter, oleuropein and hydroxytyrosol were added to each well to a concentration of 10 ⁻⁶ M to 10 ⁻⁴ M. The medium containing oleuropein was changed every 3 days, and the amount of calcium deposition was measured by the following method after 15 days and 18 days. As a control, an experiment using the aforementioned apigenin was performed.

  The cells were washed with PBS, 1 ml of 2 N HCl solution was added to each well, and the calcium phosphate salt deposited was gently agitated overnight to decalcify. The liberated calcium ion was measured using a Calcium C kit manufactured and sold by Wako Pure Chemical Industries.

  The results are shown in “FIGS. 5A, 5B, and 5C”. The vertical axis represents the amount of calcium deposition. It was found that the osteoblasts on the 15th and 18th day after the addition of oleuropein and hydroxytyrosol had a higher amount of calcium deposition than the control. It was also shown that the amount of calcium deposition increased depending on the concentration of oleuropein and hydroxytyrosol. On the other hand, it was found that the amount of calcium deposition decreased in an osteoblast to which apigenin was added in a concentration-dependent manner.

  From the above results, it was shown that oleuropein and hydroxytyrosol promoted calcification in osteoblasts in a concentration-dependent manner. However, apigenin did not act to promote osteoblast differentiation. This strongly suggests that the effect of oleuropein and hydroxytyrosol on osteoblast calcification is not a general effect on olive-derived flavonoids but an effect specific to oleuropein and hydroxytyrosol.

<Examination of the effect of oleuropein on osteoclasts>
In bone resorption, osteoclasts are known to lyse bone tissue and play an important role in bone destruction. Therefore, in order to know the effect of oleuropein etc. on osteoclasts, in vitro experiments using cultured osteoclasts were performed. A method for separating osteoclast precursor cells (spleen cells) and a method for inducing differentiation of osteoclasts are shown below.

As animals, ddY mice (male, 6 weeks old) were used. To the spleen cell fraction of the mouse, 10 mM Tris-HCl (ph7.4) containing 0.83% ammonium chloride was added to remove blood cells. The spleen-derived cells thus isolated are seeded on a 96-well plate at 2.4 × 10 ⁵ cells / well, and 50 ng / ml human sRANKL and 30 ng / ml M-CSF involved in osteoclast differentiation are added. Added and cultured for 7 days. The medium containing cytokines was changed every 3 days.

In order to know the effect of oleuropein etc. on osteoclast differentiation, TRAP staining was performed. Osteoclasts are seeded in a 96-well plate at 2.4x10 ⁵ cells / well, added with 50 ng / ml human sRANKL and 30 ng / ml M-CSF, which are involved in osteoclast differentiation, and cultured for 7 days. did. At the same time, oleuropein, hydroxytyrosol and tyrosol were added to each well to a concentration of 10 ⁻⁵ M to 10 ⁻⁴ M. The medium containing oleuropein and its derivatives was changed every 3 days, and TRAP staining was performed by the following method on the 7th day. As a control, an experiment was conducted in which the aforementioned apigenin was added.

  For TRAP staining, after fixing the cells with 3.6% formaldehyde, the cells were washed with 50 mM sodium acetate buffer, 40 mM potassium sodium tartrate buffer (pH 5.0), 0.01% naphthol AS-MX phosphate, 0.03% fast red violet LB salt. Stained for 20 minutes. After removing the staining solution, the cells were washed with distilled water, and TRAP-positive multinucleated cells were observed under an optical microscope (Olympus CK40).

  The results are shown in “FIGS. 6A and 6B”. The photograph shows a photograph of TRAP-stained cells. (A) Osteoclasts added with oleuropein, (B) osteoclasts added with hydroxytyrosol, (C) osteoclasts added with tyrosol. It was found that the formation of TRAP-positive cells (osteoclasts) was inhibited in cultured osteoclasts supplemented with oleuropein and its derivatives. In particular, osteoclasts supplemented with oleuropein were shown to strongly inhibit osteoclast formation.

  It was found that oleuropein and its derivatives have an effect of suppressing osteoclast formation. Among these, it was shown that oleuropein strongly suppressed the formation of osteoclasts. On the other hand, osteoclasts added with apigenin did not inhibit osteoclast formation at the same concentration as oleuropein. This strongly suggests that the effect on osteoclast formation inhibition such as oleuropein is not a general effect on olive-derived flavonoids but an effect specific to oleuropein and its derivatives.

  As mentioned above, as shown in Examples 1 to 3, the present inventors have promoted osteoblast differentiation activity as an action of oleuropein and hydroxytyrosol, while suppressing the formation of osteoclasts. It was found to have It has been clarified that oleuropein and its derivatives have the effect of promoting bone formation as a whole.

  The osteogenesis promoter according to the present invention can be suitably used as a pharmaceutical composition, a food composition or the like in order to promote bone formation.

It is a figure shown about the metabolite of oleuropein. It is a figure which shows the effect of oleuropein in the bone resorption model mouse | mouth by ovariectomy. The bone density of the cancellous bone at a portion 1.8 mm inside from the growth plate was measured. “Sham”: a mouse that had undergone only open surgery without removing the ovary, “OVX”: a mouse that had just removed the ovary, “Oleuropein”: a mouse that had received oleuropein after removing the ovary, “Hydroxytyrosol”: Mice treated with hydroxytyrosol after removal of ovaries, "Tyrosol": Mice treated with tyrosol after removal of ovaries It is a figure shown about the effect of oleuropein (a), hydroxytyrosol (b), and apigenin (c) on the proliferation of osteoblasts. It is a figure shown about the effect of oleuropein (a), hydroxytyrosol (b), and apigenin (c) with respect to osteoblast differentiation. (A) It is a figure shown about the effect of oleuropein (a), hydroxytyrosol (b), and apigenin (c) with respect to calcification of osteoblasts. (A) It is a figure of the TRAP dyeing | staining image shown about the effect of an oleuropein and an oleuropein derivative with respect to osteoclast formation. (B) The number of osteoclasts formed by oleuropein and its derivatives.

Claims (8)

  An osteogenesis promoter containing oleuropein and / or hydroxytyrosol as an active ingredient.   The osteogenesis promoter according to claim 1, which has an inhibitory effect on osteoclast formation.   3. The osteogenesis promoter according to claim 2, which has an effect of promoting calcification of osteoblasts.   4. The osteogenesis promoter according to claim 3, which has an osteoblast differentiation promoting action.   A pharmaceutical composition comprising the osteogenesis promoter according to claim 4 as an active ingredient.   A food composition that contains oleuropein and / or hydroxytyrosol and indicates that it is used to promote bone formation.   The bone formation promoter according to claim 1, wherein the oleuropein and / or hydroxytyrosol is derived from an olive extract. Use of the oleuropein and / or hydroxytyrosol for producing an osteogenesis promoter.