JP2000247896A - Bone quantity reduction-inhibitory composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a bone quantity reduction-inhibitory composition that has the effect to inhibit the bone quantity reduction and is useful for prevention and treatment of a variety of osteopathy, for example, osteoporosis and the like by using extract of pomegranates as an active ingredient. SOLUTION: This bone quantity reduction preventive composition includes an extract of pomegranate (preferably pomegranates grown in Iran as an active ingredient). The extract is preferably prepared by extracting it from fruits of pomegranate with a hydrophilic organic solvent, for example, ethanol, acetone or the like), polyhydric alcohol, for example, glycerol, propylene glycol or the like), water or the like. The daily dose of this composition is 50-100 mg/adult calculated as dried powder in several portions for attaining the bone quantity reduction effect. This extract is admixed to medicines, foods, beverages and feeds and they are imparted the action to inhibit the bone quantity reduction.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a composition for suppressing bone loss, comprising a pomegranate fruit extract as an active ingredient. The composition of the present invention has an effect of suppressing bone loss, and is effective for prevention and treatment of various bone diseases such as osteoporosis.

[0002]

2. Description of the Related Art In recent years, the number of people suffering from various bone diseases such as osteoporosis, fracture, rheumatism, arthritis and back pain has been rapidly increasing with the aging society, and prevention and treatment methods thereof are urgently required. . Osteoporosis and other bone disorders are caused by insufficient calcium intake, decreased calcium absorption capacity, and postmenopausal hormones.
It is said that the cause is imbalance. In order to prevent various bone diseases such as osteoporosis due to such aging, obtain as much bone mass as possible in childhood and adolescence,
Increasing the maximum bone mass is considered to be extremely important. However, since the bone mass reaches its maximum in the twenties and continues to decrease thereafter, maintaining the bone mass after middle age will be a future task. In particular, the bone loss of women after menopause is remarkable, and development of foods, medicines and feeds capable of suppressing the bone loss has been desired. On the other hand, pomegranate is native to the Asia Minor region, and is a deciduous tree that was introduced in Japan during the Heian period for medicinal or ornamental use. Pomegranate is used as a pest control and gargle. As for pomegranate, it is disclosed in Japanese Examined Patent Publication No. 5-320063 that bark and bark have antioxidant activity, but it has not been known to have an effect of suppressing bone loss.

[0003]

SUMMARY OF THE INVENTION An object of the present invention is to provide a composition for inhibiting bone loss, which has an effect of inhibiting bone loss and is effective for preventing or treating various bone diseases such as osteoporosis.

[0004]

Means for Solving the Problems The present inventors have conducted various searches for natural products that suppress the decrease in bone mass, and as a result, have found that pomegranate fruit extract suppresses the decrease in bone mass, and completed the present invention. I came to. That is, the feature of the composition for suppressing bone loss according to the present invention is that a pomegranate fruit extract is used as an active ingredient.

BEST MODE FOR CARRYING OUT THE INVENTION The pomegranate fruit which can be used in the present invention may be any pomegranate collected from Iran, the United States, Persia, etc. It is easy to use because there are many edible parts.

The pomegranate extract, which is an active ingredient of the composition for inhibiting bone loss according to the present invention, is obtained from a pomegranate fruit as a raw material, for example, a hydrophilic organic solvent such as ethanol, methanol or acetone, glycerin, 1,3, or the like. -Use of a polyhydric alcohol such as butylene glycol or propylene glycol, or a mixed solution of the above polyhydric alcohol and water, or an extract obtained by solvent extraction using water as an extraction solvent, as a pomegranate fruit extract. Can be. In particular,
It is preferable to use an organic solvent as the extraction solvent because an extract having a high content of the active ingredient can be obtained. Further, the extract obtained by the above solvent extraction may be dried by a method such as freeze-drying to obtain a powder, which can be used as a pomegranate fruit extract. Regarding the composition for inhibiting bone loss using the pomegranate fruit extract of the present invention as an active ingredient, an adult can obtain the effect of inhibiting bone loss by dividing into 50 to 100 mg per adult per day in several portions in terms of dry powder. It is effective for prevention and treatment of various bone diseases such as osteoporosis.

[0006] The composition for inhibiting bone loss of the present invention is prepared by using a composition component usually used for pharmaceutical preparations, for example, an excipient such as a bulking agent, a diluent, a solvent or a filler, a solubilizing agent, Solubilizer, emulsifier, suspending agent, dispersant, binder, lubricant,
Mix with adjuvants such as coating agents and sustained release agents, or additives such as antioxidants, preservatives, brighteners, sweeteners, coloring agents, flavoring agents, etc. Preparations in various dosage forms such as preparations, granules, pills, drinks, syrups, and liquids.

[0007] The present invention also provides a pomegranate fruit extract having an effect of suppressing bone loss, as a food material, dairy foods such as cheese, butter, fermented milk, milk drinks, drink yogurt,
It can be added to beverages such as coffee drinks and fruit juices, confectionery such as jelly, pudding, cookies, biscuits and wafers, and various foods and drinks such as frozen foods to impart a bone loss suppressing effect. The amount of the pomegranate fruit extract that can be added to these foods and drinks is not particularly limited, but the content of the pomegranate fruit extract relative to the whole food and drink is 0.01 to 10% by weight in terms of dry powder. It is preferable to mix them. Furthermore, the pomegranate fruit extract having the effect of suppressing bone loss can be added to various feeds to impart the effect of suppressing bone loss. The amount of the pomegranate fruit extract that can be blended in the feed is not particularly limited, but the pomegranate fruit extract is blended so that the content of the pomegranate fruit extract in the whole feed becomes 0.01 to 10% by weight in terms of dry powder. Is preferred. Hereinafter, the present invention will be described in detail with reference to Examples and Test Examples.

[0008]

Example 1 (Preparation of pomegranate fruit extract) 10 g of pomegranate fruit (fresh fruit) was coarsely crushed with a mixer, and acetone 30
0 ml was added, and the mixture was left for 5 days with occasional stirring. This was filtered and freeze-dried to obtain a powder.

[0009]

Example 2 (Preparation of pomegranate fruit extract) 10 g of pomegranate fruit (fresh fruit) was coarsely crushed with a mixer, 300 ml of a 50% aqueous methanol solution was added, and the mixture was left for 5 days with occasional stirring. This was filtered and freeze-dried to obtain a powder.

[0010]

Example 3 (Preparation of pomegranate fruit extract) 10 g of pomegranate fruit (raw fruit) was coarsely crushed with a mixer, and purified water 300 g
The mixture was heated for 3 hours. This was allowed to cool, filtered, and lyophilized to obtain a powder.

[0011]

Example 4 (Preparation of pomegranate fruit extract) 10 g of pomegranate fruit (raw fruit) was roughly crushed with a mixer, and ethanol
After adding 00 ml and stirring occasionally, the mixture was left for 5 days. This was filtered and freeze-dried to obtain a powder.

[0012]

Example 5 (Preparation of pomegranate fruit extract) 10 g of pomegranate fruit (raw fruit) was roughly crushed with a mixer, 300 ml of a 10% aqueous ethanol solution was added, and the mixture was left for 5 days with occasional stirring. This was filtered and freeze-dried to obtain a powder.

[0013]

Example 6 (Preparation of pomegranate fruit extract) 300 ml of acetone was added to 5 g of a freeze-dried pomegranate fruit, and the mixture was left for 5 days with occasional stirring. This was filtered and freeze-dried to obtain a powder.

[0014]

Example 7 (Preparation of pomegranate fruit extract) To 5 g of a freeze-dried pomegranate fruit was added 300 ml of a 50% methanol aqueous solution, and the mixture was left for 5 days with occasional stirring. This was filtered and freeze-dried to obtain a powder.

[0015]

Example 8 (Preparation of Pomegranate Fruit Extract) 300 ml of purified water was added to 5 g of a freeze-dried pomegranate fruit and heated for 3 hours. This was allowed to cool, filtered, and lyophilized to obtain a powder.

[0016]

Example 9 (Preparation of pomegranate fruit extract) 300 ml of ethanol was added to 5 g of freeze-dried pomegranate fruit, and the mixture was left for 5 days with occasional stirring. This was filtered and freeze-dried to obtain a powder.

[0017]

Example 10 (Preparation of Pomegranate Fruit Extract) 300 ml of 10% ethanol aqueous solution was added to 5 g of freeze-dried pomegranate fruit.
Was added and left for 5 days with occasional stirring. This was filtered and freeze-dried to obtain a powder. Next, test examples in which the effects of the present invention were confirmed by cell experiments and animal experiments will be described.

[0018]

[Test Example 1] Extraction of pomegranate fruit obtained in Examples 1 to 10
The effect of the bone resorption on the powder of the product was examined. That is,
Excision of long bone of ICR mouse 10-20 days old
After removing soft tissue, α-M containing 5% fetal bovine serum
Mechanically mince bone in EM solution, whole bone including osteoclasts
Medullary cells were obtained. About 2 × 10 cells on ivory pieces
⁶Α-M containing 5% fetal calf serum to give a cell mass
Spot with EM solution. After two hours, extract each pomegranate fruit
Was added to a final concentration of 10 μg / ml.
Α-MEM solution containing 5% fetal bovine serum_Two
Cultured at 37 ° C for 5 days in the presence of osteoclasts to resorb bone
Was examined. After culturing, peel the cells from the ivory pieces
Syrin staining and image analysis (PIASLA-555,
Image analysis by PIAS) and bone resorption fossa (pit)
The number was measured. From the measurement results, the bone resorption defined by the following equation
The activity (%) was determined, and the effect of preventing bone resorption was evaluated.

[0019]

## EQU1 ## Bone resorption activity (%) = (number of bone resorption pits in added group / number of bone resorption pits in non-added group) × 100

The results are shown in Table 1. In the addition group in which each pomegranate fruit extract obtained in Examples 1 to 10 was added to the medium,
The bone resorption activity was suppressed to about 20 to 30%, which revealed that the pomegranate fruit extract had a bone resorption preventing action.

[0021]

[Table 1]

[0022]

Test Example 2 In 100 g of feed, 1 g each of the pomegranate fruit extract powders obtained in Examples 1, 3 and 5; casein 20 g; corn starch 15 g; cellulose 5 g;
Corn oil 5 g, sucrose 49.2 g, DL-methionine 0.
3 g, 3.5 g of a mineral mixture (“AIN-76 Mineral Mix”, manufactured by Oriental Yeast Co., Ltd.) and 1 g of a vitamin mixture (“AIN-76 Vitamin Mix”, manufactured by Oriental Yeast Co., Ltd.) to prepare a feed, The effect of suppressing bone loss was examined. As a control, a feed without pomegranate fruit extract powder was prepared, and the same test was performed. Forty 7-month-old SD female rats were grouped in groups of 8
The animals were divided into groups, and 8 animals in the sham operation group underwent sham operation, and 32 animals in the remaining 4 groups underwent ovariectomy. After a one-week recovery period, the ovariectomized operation was performed, and the experimental feed supplemented with the pomegranate fruit extract powder obtained in Example 1 was added to the 8 animals in the addition group I, and the 8 animals in the addition group II were added to the 8 animals. The experimental feed to which the pomegranate fruit extract powder obtained in Example 3 was added, and the experimental feed to which the pomegranate fruit extract powder obtained in Example 5 was added were supplied to eight animals of the addition group III, respectively. They were bred for weeks. Eight animals in the sham operation group and eight animals in the non-addition group were fed with pomegranate fruit extract powder-free feed as a control and bred for 4 weeks. Four weeks later, the femur was excised, and the bone density was measured by the DEXA method using a bone density measurement device (Dichroma Scan DCS-600A, manufactured by Aloka). Table 2 shows the measurement results. In the non-addition group, the bone density was significantly lower than in the sham operation group, and a decrease in bone mass due to the operation was observed. On the other hand, the bone density was significantly higher in the additive group than in the non-additive group, and it was clarified that bone loss due to surgery was suppressed by ingestion of the pomegranate fruit extract. Also, almost no difference was found depending on the type of the extraction solvent.

[0023]

[Table 2] The numerical values indicate the mean value ± standard deviation. * Indicates that there is a significant difference from the non-added group (p <
0.05)

[0024]

Example 11 Production of a milk beverage containing pomegranate fruit extract powder; pomegranate fruit extract powder obtained in Example 1 was added to raw milk so as to give 1 g per 100 ml;
After homogenizing at a pressure of 0 kg / cm ² ,
For 4 seconds to produce a milk drink having an effect of suppressing bone loss.

[0025]

EXAMPLE 12 Production of yogurt formulated with pomegranate fruit extract powder; was dissolved in water to non-fat dry milk having a solid ratio of 12%, after heat sterilization at 90 ° C. 20 min, cooled to 25 ° C., L They were inoculated with .acidophilus and S.thermophilus. When the lactic acidity reached 1.0% and the pH reached 4.3, the mixture was cooled to 5 ° C. The starter culture thus obtained is inoculated into sterilized raw milk having a fat content of 5% by 5%, and 1.5 g of the pomegranate fruit extract powder obtained in Example 1 is further added.
Was added. After inoculation of the starter culture, fermentation, flavoring, and cooling were carried out in the usual manner to produce a yogurt having an effect of suppressing bone loss.

[0026]

Example 13 Production of biscuits containing pomegranate fruit extract; dough having the composition shown in Table 3 was prepared according to a conventional method.
After being molded, the biscuit was roasted to give the effect of suppressing bone loss.

[0027]

[Table 3] 粉 Flour 50.0 (% by weight) Sugar 20.0 Salt 0.5 Margarine 12.5 Egg 12.5 Water 2.5 Pomegranate obtained in Example 1 Fruit extract powder 2.0 ──────────────────────

[0028]

Example 14 Production of tablets containing pomegranate fruit extract; Tablets having the composition shown in Table 4 and having the effect of suppressing bone loss were produced using a tableting machine.

[0029]

Table 4 水 Hydrous crystalline glucose 88.5 (% by weight) Pomegranate fruit extract powder obtained in Example 1 10.0 Sugar ester 1.0 Spice 0.5 ──────────────────────

It is desirable to administer 5 to 15 tablets per day for the prevention or treatment of osteoporosis.

[0031]

Example 15 Production of a dog breeding feed containing a pomegranate fruit extract; raw materials shown in Table 5 were mixed with a cake mixer to produce a dog breeding feed (dog food) having an effect of suppressing bone loss.

[0032]

[Table 5] ────────────────────── Soybean meal 11 (wt%) skim milk powder 14 soybean oil 4 corn oil 2 palm oil 2 corn starch 30 wheat flour 15 Bran 8 Vitamin mixture 2 Mineral mixture 9 Cellulose 2 Pomegranate fruit extract powder obtained in Example 1 1──────────────────────

[0033]

Industrial Applicability The composition for inhibiting bone loss comprising the pomegranate fruit extract of the present invention as an active ingredient has an effect of inhibiting bone loss, and has various bone diseases such as osteoporosis (fracture, rheumatism,
It is effective in preventing and treating arthritis and back pain. Further, the pomegranate fruit extract is useful because it can be added to and blended with foods, pharmaceuticals, feeds, and the like, and can exert an effect of suppressing bone loss.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61P 19/10 A61K 31/00 619E F-term (Reference) 2B150 AA06 AB10 DD31 DD45 DD57 4B016 LC07 LG01 LP02 4B018 MD52 ME05 MF01 4C088 AB12 AC04 BA08 BA09 BA10 CA05 CA06 CA08 MA52 NA14 ZA96 ZA97 ZC61

Claims (3)

[Claims] 1. A composition for inhibiting bone loss, comprising a pomegranate fruit extract as an active ingredient. 2. A medicament, food or drink or feed containing a pomegranate fruit extract and having an effect of inhibiting bone loss. 3. A method for producing a pomegranate fruit extract having an effect of suppressing bone loss, comprising a step of extracting pomegranate fruit with an organic solvent, water, or a mixture of an organic solvent and water.