JP5307329B2 - Bone formation promoter containing β-cryptoxanthin as an active ingredient - Google Patents

Description

  The present invention relates to a bone formation promoter containing β-cryptoxanthin as an active ingredient, a preventive / therapeutic agent for bone diseases such as osteoporosis, a functional food for the prevention / treatment of bone diseases such as osteoporosis to which β-cryptoxanthin is added, or The present invention relates to a method for screening an active ingredient for promoting osteogenesis or preventing / treating bone diseases using β-cryptoxanthin as a lead compound.

  It is considered that various bone diseases occur due to a decrease in the amount of calcium in bone and the like due to bone metabolism and bone dysplasia. Representative bone diseases include fractures, osteomalacia, osteopenia, osteoporosis, and back pain. Among these bone diseases, osteoporosis, in particular, is caused by the loss of bone mass due to the relative superiority of bone resorption due to the imbalance between bone resorption and bone formation due to aging. It shows a pathological condition in which the strength of the bones decreases and fractures are likely to occur. Especially in women, bone loss rapidly occurs due to menopause and ovariectomy. Osteoporosis not only causes fractures and severe pain, but also causes bedridden elderly people in particular, so effective treatment is required from the viewpoint of improving the quality of life in an aging society. . Since osteoporosis is difficult to treat after it develops, it is important to make efforts to prevent it, and it is essential to increase bone mass from a young age. It has become deeply recognized that foods that promote formation must be actively consumed. Currently, calcium, magnesium and vitamin D are mainly used as foods for strengthening bones. Casein phosphopeptides that promote absorption of calcium from the intestinal tract have also been used.

  As therapeutic agents for bone diseases such as osteoporosis, active vitamin D3, female hormone (estrogen), calcitonin, and ipriflavones are used clinically. Recently, osteoclasts of polyisoprenoid derivatives represented by vitamin K2 are used. An anti-osteoporosis agent (Japanese Patent Laid-Open No. 7-215849) based on a formation inhibitory action has also been developed. Further, a bone strengthening and anti-osteoporosis composition comprising, as main active ingredients, a bone strengthening agent containing casein phosphopeptide and genistein (Japanese Patent Laid-Open No. 2001-302539), saponin, daidzin, daidzein, genistin and genistein ( JP-A-2000-191526), a bone mass-promoting composition that exhibits an anti-osteoporosis effect using a wasabi extract as an active ingredient (JP-A-10-279492), and a bone disease therapeutic agent that contains zinc acetamate as an active ingredient (Japanese Patent Laid-Open No. 10-218767), composition for promoting bone formation and preventing bone mineral loss (isolated from Japanese Patent Laid-Open No. 10-114653) containing isoflavone as a main active ingredient, anti-osteoporosis composition strengthened with both vitamin K2 and zinc An article (Japanese Patent Laid-Open No. 10-36256) is also known.

  On the other hand, β-cryptoxanthine (β-cryptoxanthine; molecular weight 552) is known as an ethanol-soluble carotenoid, and is predominantly contained in citrus fruits in Wenzhou mandarin orange and 1-2 mg in one fruit. This β-cryptoxanthin not only has the characteristics of provitamin A as a nutritional component, but in recent research on anticancer substances, β-carotene is a carotenoid contained in green-yellow vegetables such as carrots. It has become clear that it has a higher anticancer effect than that (Biol. Pharm. Bull. 18, 2, 227, 1995). Since β-cryptoxanthin is important as an anti-carcinogenic component in this way, it has a high quality with a β-cryptoxanthin content comparable to Wenzhou mandarin orange to help develop citrus strains and citrus processed foods that are enriched with β-cryptoxanthin. Production of citrus fruits, isolation of genes that synthesize β-cryptoxanthin (JP-A-11-155777, JP-A-11-46770), development of technology for separating β-cryptoxanthin from citrus in large quantities, etc. Has been done.

  In addition, the separation method of β-cryptoxanthin from citrus fruits such as Unshu mandarin orange is well known (Okayama University Journal of Agriculture, 69, 17-25, 1987, Bulletin of Tokyo Medical University 18, 1-7, 1992, Journal of Food Biochemistry., 18, 273-283, 1995). Recently, a solvent extract containing β-cryptoxanthin was obtained from a raw material precipitate obtained by squeezing mandarin orange juice, and this was hydrolyzed. The degradation product was introduced into a first column packed with silica powder having an average particle size of 10 to 80 μm together with the primary developing solvent at a flow rate of 2 cm / min or more to separate and remove the fraction containing β-cryptoxanthin. Later, the separated product was introduced into a second column packed with octadecylsilane silica having an average particle size of 10 to 80 μm together with a secondary developing solvent at a flow rate of 2 cm / min or more, and 95 wt% β-cryptoxanthin was added. More than A method for producing high-purity β-cryptoxanthin that separates the fractions contained in the product (Japanese Patent Laid-Open No. 2000-136181), and citrus fruits after squeezing, filtering or sieving, and then centrifuging the precipitate into the enzyme A method for producing a carotenoid-rich pulp, characterized by adding an agent, freezing, thawing, and dehydrating, repeating the operation of adding and dehydrating the carotenoid-rich pulp, drying and grinding, A carotenoid-rich pulp with enhanced content of β-cryptoxanthin and the like and a method for producing the powder (Japanese Patent Laid-Open No. 2000-23637) have been proposed.

  Furthermore, as a method for rapidly searching for potential analogs of lead compounds, radicals are temporarily bonded to bulky space-retaining groups, and radical 3D models are registered in a combinatorial ghost database. For any molecular structure accessible in the database, detect any atom that exhibits the characteristics of a pharmacophore-type physical property; for each pharmacophore pair detected in each molecular structure, relate to the total conformation of this molecule All distances between atoms to calculate and create a distance distribution density; create a three-dimensional fingerprint vector that includes all distance distribution densities of a pharmacophore pair; explain the relative importance of pharmacophore features Define a scoring function for each molecular fingerprint; The above scoring function that maximizes lead compounds can compare these fingerprints with each fingerprint in the potential library, and scoring functions can give score values below a certain threshold A method for preparing and screening a chemically possible combinatorial product from a large database of 3D multi-dimensional structure fingerprints by searching for molecules in a specific library is known (Japanese Patent Publication No. 2001-521943). ).

  The eight therapeutic agents currently approved in Japan for bone diseases represented by osteoporosis are all bone resorption inhibitors (suppresses the dissolution of bone), and the only mevalonic acid synthesis inhibitor, statin. Although reported to have a formation promoting action, it was a knowledge at the gene level, and in fact, the bone formation promoting effect was weak. An object of the present invention is to provide a bone formation promoter that has a prominent effect capable of actively promoting bone formation to prevent and treat bone diseases, and has both a bone formation promoting action and a bone resorption inhibiting action. Providing preventive / therapeutic agents for bone diseases such as osteoporosis, screening methods for active ingredients for the prevention and treatment of bone diseases using compounds having both osteogenesis promoting and bone resorption inhibiting effects as lead compounds There is.

  The present inventors have found that β-cryptoxanthin, which is abundant in the epidermis and pulp of Satsuma mandarin orange, has a bone formation promoting effect and a bone disease prevention / treatment effect. That is, the femoral bone metaphysis and metaphyseal tissue are cultured in a culture solution containing β-cryptoxanthin, and the amount of calcium in the bone tissue, the expression level of the bone mineralization promoting enzyme, and the cell number index in the bone tissue In both cases, a significant increase was confirmed, and β-cryptoxanthin promoted protein synthesis in the cancellous bone (metaphyseal tissue) and cortical bone (diaphyseal tissue) of the femoral tissue. , Found to increase bone formation. In addition, bone tissue is cultured in the coexistence of parathyroid hormone (PTH) and β-cryptoxanthin, which has a bone mineral dissolving (bone resorption) action and plays a physiological role in the pathogenesis of osteoporosis due to aging. It was also confirmed that a decrease in calcium content in the metaphyseal tissue can be significantly suppressed. Furthermore, when β-cryptoxanthin was orally administered to rats, the amount of calcium in the diaphysis and metaphyseal tissue, the expression level of bone mineralization promoting enzyme, and the amount of DNA serving as a cell number index in bone tissue A significant increase was observed, and it was confirmed that oral administration of β-cryptoxanthin produced a bone mass enhancement effect. From these results, it was clarified that β-cryptoxanthin enhances bone formation and suppresses bone resorption, thereby exerting an effect of maintaining and increasing bone mineral content and functioning as an anti-osteoporosis factor. . And the effect confirmed by such a tissue culture system has been confirmed empirically to be almost 100% effective even in an oral experiment. The present invention has been completed based on these findings.

That is, the present invention relates to (1) an osteogenesis promoter (except for a prophylactic / therapeutic agent for bone diseases) , characterized by comprising β-cryptoxanthin as an active ingredient.

The present invention also provides (2) an osteogenesis promoter (except for a prophylactic / therapeutic agent for bone diseases) characterized by comprising a β-cryptoxanthin-containing composition as an active ingredient, and (3) β-crypt. The xanthine-containing composition relates to an osteogenesis promoter (excluding prophylactic / therapeutic agents for bone diseases) according to (2) above, wherein the xanthine-containing composition is a processed product of Unshu oranges .

  According to the present invention, by using β-cryptoxanthin as an active ingredient, an osteogenesis promoter having a remarkable effect capable of actively promoting osteogenesis and preventing / treating bone disease, and osteogenesis promotion It is possible to provide a prophylactic / therapeutic agent for bone diseases such as osteoporosis having both an action and an action of suppressing bone resorption.

  The osteogenesis promoter of the present invention is not particularly limited as long as it contains β-cryptoxanthin or a β-cryptoxanthin-containing composition as an active ingredient, and prevention / treatment of the bone disease of the present invention. The drug is not particularly limited as long as it contains β-cryptoxanthin or a β-cryptoxanthin-containing composition as an active ingredient, and the functional food or food for preventing or treating the bone disease of the present invention. The material is particularly limited as long as it is a food or food material having a bone disease prevention / treatment function, which is used for the prevention / treatment of bone disease to which β-cryptoxanthin or a composition containing β-cryptoxanthin is added. In addition, as the composition for feed of the present invention, any composition containing a β-cryptoxanthin or a β-cryptoxanthin-containing composition may be used. The bone diseases may include fractures, osteomalacia, osteopenia, osteoporosis, back pain, etc. Among them, postmenopausal osteoporosis, estrogen-deficient osteoporosis, senile osteoporosis, steroid-induced osteoporosis Suitable examples thereof include metabolic bone diseases such as osteoporosis and osteomalacia.

  The method for producing β-cryptoxanthin and β-cryptoxanthin-containing compositions is not particularly limited, including known methods such as a method of extracting and generating from citrus fruits, and a method of using a gene encoding a β-cryptoxanthin producing enzyme. As the supply source, it is preferable to use Unshu mandarin, which contains 1 to 2 mg per piece, and contains β-cryptoxanthin more than 60 times that of other citrus fruits such as oranges, grapefruits and lemons. (Flaved) About 8 mg per 100 g and about 1 mg of β-cryptoxanthin per 100 g of fruit juice Sugiyama Wenzhou and other high β-cryptoxanthin-containing varieties Wenzhou oranges and high β-crypts produced by crossing with Wenzhou oranges It is preferable to use a xanthine-containing variety. Here, the β-cryptoxanthin-containing composition refers to a mixture of β-cryptoxanthin in which the β-cryptoxanthin content is artificially increased. Moreover, it does not restrict | limit especially as a method of processing a Satsuma mandarin and obtaining a beta-cryptoxanthin containing composition, For example, in addition to the above-mentioned patent documents 10 and patent documents 11, non-patent literature 2-4 processing methods are mentioned. Can be mentioned.

  When using β-cryptoxanthin or a β-cryptoxanthin-containing composition as a pharmaceutical agent such as a bone disease preventive / therapeutic agent, pharmaceutically acceptable ordinary carriers, binders, stabilizers, excipients, Various preparation ingredients such as a diluent, a pH buffer, a disintegrant, a solubilizer, a solubilizer, and an isotonic agent can be added. In addition to these, the above-described known bone formation promoting action and / or bone resorption suppressing action, and minerals such as calcium, magnesium, and phosphorus can be used in combination. These preventive or therapeutic agents can be administered orally or parenterally. That is, it can be administered orally in commonly used dosage forms, such as powders, granules, capsules, syrups, suspensions, etc., or, for example, in dosage forms such as solutions, emulsions, suspensions, etc. These can be administered parenterally in the form of injections or can be administered intranasally in the form of sprays, but are preferably administered orally. The dose can be appropriately selected depending on the purpose of prevention or treatment, the type and severity of the bone disease, the age of the patient, and the like.

  There are no particular restrictions on the type of food or food material that has a bone disease prevention / treatment function, which is used for the prevention / treatment of bone disease, to which β-cryptoxanthin or a β-cryptoxanthin-containing composition is added. Yogurt, drink yogurt, juice, milk, soy milk, liquor, coffee, tea, sencha, oolong tea, sports drinks and other beverages, baked confectionery such as pudding, cookies, bread, cakes, jelly, rice crackers, Japanese confectionery such as sheep candy, Bread and confectionery such as frozen desserts and chewing gum, noodles such as udon and soba, fish paste products such as kamaboko, ham and fish sausage, seasonings such as miso, soy sauce, dressing, mayonnaise, sweeteners, cheese, butter Dairy products such as tofu, konnyaku, other boiled rice cakes, dumplings, croquettes, salads etc. The These foods and food materials may be used in combination with the above-mentioned known bone formation promoting action and / or bone resorption inhibiting action, and minerals such as calcium, magnesium and phosphorus.

  Feed composition containing β-cryptoxanthin or β-cryptoxanthin-containing composition is advantageously used for breeding livestock and poultry such as pigs, cattle, chickens, pets such as dogs and cats, and cultured seafood The feed composition can include known substances such as the above-mentioned ipriflavones for promoting bone formation and / or inhibiting bone resorption, calcium, magnesium, phosphorus, iron, zinc, manganese, copper, etc. Can be used together.

  The method for screening active ingredients for the prevention and treatment of bone diseases such as osteoporosis of the present invention is not particularly limited as long as it is a screening method using β-cryptoxanthin as a lead compound. Such β-crypts are not limited. The screening method using xanthine as a lead compound makes it possible to develop a more effective osteogenesis promoter and a bone disease prevention / treatment agent. In order to screen for an osteogenesis promoter or a bone disease prevention / treatment agent using β-cryptoxanthin as a lead compound, for example, a combinatorial chemistry method such as the method described in Patent Document 12 described above can be used efficiently. It can be carried out. In addition, without using combinatorial chemistry methods, it is possible to screen for osteogenesis promoters and preventive / therapeutic agents for bone diseases using β-cryptoxanthin as a lead compound using classical structure-activity relationship techniques. . And the bone formation promoter and bone disease prevention / treatment agent obtained by the screening method using β-cryptoxanthin as a lead compound are also included in the present invention.

  EXAMPLES Hereinafter, although an Example demonstrates this invention more concretely, the technical scope of this invention is not limited to these illustrations.

[Method]
(Culture of rat bone tissue piece)
Rats (Wistar male: 4-5 weeks old) (purchased from Japan SLC Co., Ltd., solid feed Oriental yeast (MF)) were aseptically removed from the femur under ether anesthesia, and the femur was 0.25 M After washing with sucrose solution, it is divided into diaphyseal part (cortical bone) and metaphyseal part (cancellous bone), and contains β-cryptoxanthin etc. in, and cultured in the bone 48 hours tissue piece under conditions of 37 ° _C., in an incubator of 5% CO 2 -95% air. In addition, β-cryptoxanthin is “β-cryptoxanthin” manufactured by Extrasynthes, cycloheximide is “cycloheximide” manufactured by Sigma, β-carotene is “β-carotene” manufactured by Sigma, xanthine is the sigma "xanthine", parathyroid hormone to "parathyroid hormone" sigma, prostaglandin E ₂ was used sigma "prostaglandin E _2". In addition, the case where it culture | cultivated only with a culture solution, without adding beta-cryptoxanthin etc. was set as the control | contrast.

(Measurement of bone calcium)
The amount of calcium in the bone tissue was measured. After culturing in an incubator, the tissue pieces were washed with a 0.25M sucrose solution, dried, and then the bone weight was measured. Thereafter, concentrated nitric acid was added to the tissue pieces to make them incinerate at 120 ° C. for 12 hours, and the amount of bone calcium was quantified using an atomic absorption spectrophotometer (“Perkin Elmer 303” manufactured by Perkin Elmer).

(Measurement of alkaline phosphatase activity)
The expression level of alkaline phosphatase, the most important enzyme for promoting bone mineralization, was examined. After culturing in an incubator, the tissue pieces were washed with 0.25 M sucrose solution, crushed in 3 ml of 6.5 mM barbital buffer (pH 7.4), and sonicated. This solution was centrifuged, and the supernatant was measured as an enzyme solution according to the method of Walter and Schutt (in Method of Enzymatic Analysis, Vol1-2, p856, Academic Press, New York, 1965). Specifically, 0.05 ml of enzyme solution was added to 2 ml of diethanolamine buffer (pH 9.8) using p-nitrophenyl phosphate as a substrate, incubated at 37 ° C. for 30 minutes, and 10 ml of 0.05N NaOH was added. Absorbance (405 nm) was measured using a photometer, and the bone alkaline phosphatase activity of a therapeutic agent for bone and a compound known to act on bone was examined.

(Quantification of DNA content)
The amount of DNA was quantified as an index of the number of cells in bone tissue. After culturing in an incubator, the tissue pieces were washed with a 0.25M sucrose solution and the wet weight was measured. Then, it grind | pulverized in 4 ml of 0.1N NaOH, and it was made to osmose | permeate at 4 degreeC for 24 hours. This solution was centrifuged, and the supernatant was used as a sample and quantified according to the method of Ceriotti et al. (J. Biol. Chem., 241: 34-77, 1951). That is, 1 ml of concentrated hydrochloric acid and 1 ml of 0.04% indole solution were added to 2 ml of sample, heated to 100 ° C. in boiling water, rapidly cooled, extracted with 4 ml of chloroform, the chloroform layer was collected, and a spectrophotometer ( 490 nm) was used to measure the amount of DNA in the bone.

(Oral administration of β-cryptoxanthin to rats)
Rats (Wistar males; 4-5 weeks old) were treated with 3 kinds of β-cryptoxanthin solutions (10, 25 and 50 μg / ml corn oil) in which β-cryptoxanthin was dissolved in corn oil. Was administered orally once a day for 100 g of rat body weight for 7 days. At 24 hours after the final administration, the femur was removed, muscles and the like were removed, and the bone component was measured by dividing it into a diaphysis and a metaphysis.

[result]
(Expression of action of β-cryptoxanthin to promote osteogenesis)
The bone component increasing effect of β-cryptoxanthin was examined. In the culture medium containing β-cryptoxanthin (10 ⁻⁸ to 10 ⁻⁵ M), the diaphysis and metaphysis of the femur were cultured for 48 hours. The amount of calcium in bone tissue, alkaline phosphatase curative (bone mineralization promoting enzyme) and deoxyribonucleic acid (DNA; index of the number of cells in bone tissue) were measured by the method described in Example 1. The results are shown in Table 1 and FIGS. Each test group was measured 6 to 8 times, and indicated by an average value and a standard error. In addition, a significant difference test is performed using Student's t-test. If the P value is 0.01 or less (**) or 0.05 or less (*) compared to the control, there is a statistically significant difference. It was. As a result, cryptoxanthin (10 ⁻⁸ to 10 ⁻⁵ M) caused a significant increase in the amount of calcium in the diaphysis and metaphyseal tissue. Cryptoxanthine (10 ⁻⁸ to 10 ⁻⁵ M) significantly increases the diaphyseal alkaline phosphatase activity, and cryptoxanthin (10 ⁻⁷ to 10 ⁻⁵ M) increases the enzyme activity at the metaphysis. Caused. Furthermore, the amount of diaphyseal tissue and metaphyseal tissue DNA was significantly increased in the presence of cryptoxanthin (10 ⁻⁷ to 10 ⁻⁵ M).

(Influence of protein synthesis inhibitors)
The effect of protein synthesis inhibitors on the expression of the bone component increasing effect of β-cryptoxanthin was investigated. As the protein synthesis inhibitor, cycloheximide that acts on 60S ribosomes of eukaryotic cells and inhibits transfer reaction in peptide chain elongation was used. In the presence of β-cryptoxanthin (10 ⁻⁶ M), in the presence of cycloheximide (10 ⁻⁶ M), in the presence of β-cryptoxanthin (10 ⁻⁶ M) and cycloheximide (10 ⁻⁶ M) , Calcium content, alkaline phosphatase activity and DNA content in bone tissue were measured. The results are shown in Table 2 and FIGS. Each test group was measured 6 times, and the average value and standard error were shown. In addition, a significant difference test was performed using Student's t-test. If the P value was 0.01 or less (*) as compared with the control, the difference was statistically significant. Calcium content, bone alkaline phosphatase activity and bone DNA content in the diaphysis and metaphysis tissues increased in the presence of β-cryptoxanthin (10 ⁻⁶ M) were significant in the presence of cycloheximide (10 ⁻⁶ M). Declined. These results show that β-cryptoxanthin increases bone formation by enhancing its protein synthesis in cancellous bone (metaphyseal tissue) and cortical bone (diaphyseal tissue) of femoral tissue. It is revealed.

(Comparative example)
Instead of β-cryptoxanthin, the effects of β-carotene and xantine (2,6-dihydroxypurine) on the amount of bone calcium were examined. 10 ⁻⁷ M of β-carotene and 10 ⁻⁶ M of xanthine were present in the culture solution, respectively, and the diaphyseal part or metaphyseal tissue was cultured for 48 hours by the method described in Example 1; The amount of calcium was measured. The results are shown in Table 3 and FIGS. 13 and 14. Each value in Table 3 shows the mean value ± standard error of 6-8 rats. As is apparent from Table 3, β-carotene and xanthine did not exhibit a significant increase in the amount of calcium in bone tissue.

(Expression of inhibitory action of β-cryptoxanthin on bone resorption)
The inhibitory effect of β-cryptoxanthin on bone mineral dissolution was examined. Parathyroid hormone (PTH) is a peptide hormone that is secreted from the parathyroid gland and exerts a bone mineral dissolving (bone resorption) action, and plays a pathophysiological role in the development of osteoporosis associated with aging. Similarly, it is known that prostaglandin E ₂ also causes a physiological bone mineral dissolution. Therefore, after culturing the diaphyseal and metaphyseal tissues of the femur for 48 hours in the presence of 10 ⁻⁷ M PTH or 10 ⁻⁵ M prostaglandin E ₂ , the amount of calcium in the bone tissue Was measured. Moreover, after culturing similarly in the presence of these and β-cryptoxanthin (10 ⁻⁸ to 10 ⁻⁶ M), the amount of calcium in the bone tissue was measured. The results are shown in Table 4 and FIGS. Each test group was measured 6 to 8 times, and indicated as an average value and standard error. Further, determined using the Student's t-test for significant difference test, P value of 0.01 or less as compared to the control (*), or P value compared to PTH and prostaglandin E ₂ alone 0. If it is 01 or less (#), it is considered that there is a statistically significant difference. When bone tissue was cultured in the presence of PTH, the amount of calcium in the diaphysis and metaphyseal tissue was significantly reduced. This decrease was significantly suppressed in the presence of β-cryptoxanthin (10 ⁻⁸ to 10 ⁻⁶ M). Similarly, prostaglandin E ₂ (10 ⁻⁵ M), which causes physiological bone mineral dissolution, caused a significant decrease in the amount of calcium in bone tissue, but this decrease was caused by β-cryptoxanthin ( 10 ^-8 to 10 ^-6 M).

  Based on the above results, it was confirmed that β-cryptoxanthin promotes and promotes bone formation and suppresses bone resorption, thereby maintaining and increasing bone mineral content and functions as an anti-osteoporosis factor. It was.

(Increase of bone component by oral administration of β-cryptoxanthin to rats)
It was examined whether or not bone components increased when β-cryptoxanthin was orally administered to rats. After administration of β-cryptoxanthin for 7 days (10, 25, 50 μg / 100 g body weight / day), the amount of calcium in bone tissue, alkaline phosphatase cure (bone mineralization promoting enzyme), and deoxyribonucleic acid (DNA; in bone tissue) The number of cells) was measured by the method described in Example 1. The results are shown in Table 5 and FIGS. In addition, each test group measured 6 times each, and showed with the average value and the standard error. In addition, a significant difference test was performed using Student's t-test. If the P value was 0.01 or less (*) as compared with the control, the difference was statistically significant. As a result, the amount of calcium in the diaphysis and metaphyseal tissue was significantly increased by the administration of β-cryptoxanthin (10, 25, 50 μg / 100 g body weight / day). Alkaline phosphatase activity (bone mineralization promoting enzyme) in the diaphysis and metaphysis tissue was significantly increased by the administration of β-cryptoxanthin (10, 25, 50 μg / 100 g body weight / day). The amount of DNA in bone tissue (an index of the number of cells in bone tissue) is significantly increased by administration of β-cryptoxanthin (25 and 50 μg / 100 g body weight / day) in the diaphysis, and in the metaphysis, β -Significant increase with administration of cryptoxanthin (10, 25, 50 μg / 100 g body weight / day).

  From the above, it was confirmed that β-cryptoxanthin increases the amount of bone components and has a bone mass promoting effect by oral administration. Therefore, β-cryptoxanthin is useful as an osteogenesis promoter and is considered effective for the prevention and treatment of osteoporosis.

It is a figure which shows the measurement result of the amount of bone calcium in the diaphyseal structure | tissue of (beta) -cryptoxanthin of this invention. It is a figure which shows the measurement result of the bone alkaline phosphatase activity in the shaft tissue of the beta-cryptoxanthin of this invention. It is a figure which shows the measurement result of the amount of bone DNA in the diaphyseal structure | tissue of (beta) -cryptoxanthin of this invention. It is a figure which shows the measurement result of the amount of bone calcium in the metaphysis part structure | tissue of (beta) -cryptoxanthin of this invention. It is a figure which shows the measurement result of the bone alkaline phosphatase activity in the metaphysis structure | tissue of (beta) -cryptoxanthin of this invention. It is a figure which shows the measurement result of the amount of bone DNA in the metaphysis tissue of (beta) -cryptoxanthin of this invention. It is a figure which shows the measurement result of the amount of bone calcium in the diaphyseal structure | tissue in the coexistence of (beta) -cryptoxanthin of this invention, and a protein synthesis inhibitor. It is a figure which shows the measurement result of the bone alkaline phosphatase activity in the diaphyseal structure | tissue in the coexistence of (beta) -cryptoxanthin of this invention, and a protein synthesis inhibitor. It is a figure which shows the measurement result of the amount of bone DNA in the diaphyseal structure | tissue in the coexistence of (beta) -cryptoxanthin and protein synthesis inhibitor of this invention. It is a figure which shows the measurement result of the amount of bone calcium in the metaphysis part structure | tissue in the coexistence of (beta) -cryptoxanthin of this invention, and a protein synthesis inhibitor. It is a figure which shows the measurement result of the bone alkaline phosphatase activity in the metaphysis tissue in the coexistence of (beta) -cryptoxanthin of this invention, and a protein synthesis inhibitor. It is a figure which shows the measurement result of the amount of bone DNA in the metaphysis tissue in the coexistence of (beta) -cryptoxanthin of this invention, and a protein synthesis inhibitor. It is a figure which shows the measurement result of the amount of bone calcium in the diaphyseal structure | tissue in coexistence of (beta) -cryptoxanthin of this invention, and (beta) -carotene, and coexistence of (beta) -cryptoxanthin and xanthine. It is a figure which shows the measurement result of the amount of bone calcium in the metaphyseal part structure | tissue in coexistence of (beta) -cryptoxanthin of this invention, and (beta) -carotene, and coexistence of (beta) -cryptoxanthin and xanthine. It is a figure which shows the measurement result of the amount of bone calcium in the diaphyseal structure | tissue in the coexistence of (beta) -cryptoxanthin of this invention, and a parathyroid hormone. It is a figure which shows the measurement result of the amount of bone calcium in the metaphyseal structure | tissue in the coexistence of (beta) -cryptoxanthin and parathyroid hormone of this invention. Is a graph showing measurement results of the bone calcium content in the diaphysis tissue in the presence of a β- cryptoxanthin and prostaglandin E ₂ of the present invention. Is a graph showing measurement results of the bone calcium content in the metaphysis tissue in the presence of a β- cryptoxanthin and prostaglandin E ₂ of the present invention. It is a figure which shows the measurement result of the amount of bone calcium in a diaphyseal tissue when the beta-cryptoxanthin of this invention is orally administered to a rat. It is a figure which shows the measurement result of the bone alkaline phosphatase activity in a diaphyseal tissue when the beta-cryptoxanthin of this invention is orally administered to a rat. It is a figure which shows the measurement result of the amount of bone DNA in a diaphyseal tissue when (beta) -cryptoxanthin of this invention is orally administered to a rat. It is a figure which shows the measurement result of the amount of bone calcium in the metaphysis tissue when the beta-cryptoxanthin of this invention is orally administered to a rat. It is a figure which shows the measurement result of the bone alkaline phosphatase activity in the metaphysis tissue when the beta-cryptoxanthin of this invention is orally administered to a rat. It is a figure which shows the measurement result of the amount of bone DNA in the metaphysis tissue when (beta) -cryptoxanthin of this invention is orally administered to a rat.

Claims (3)

An osteogenesis promoter characterized by comprising β-cryptoxanthin as an active ingredient (excluding preventive and therapeutic agents for bone diseases) . An osteogenesis promoter characterized by comprising a β-cryptoxanthin-containing composition as an active ingredient (excluding preventive and therapeutic agents for bone diseases) . The bone formation promoter according to claim 2, wherein the β-cryptoxanthin-containing composition is a processed product of Unshu mandarin orange (except for a prophylactic / therapeutic agent for bone diseases) .