JP3250071B2 - Anti-osteoporosis composition - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to an anti-osteoporosis composition fortifying both vitamin K and soy isoflavone.

[0002]

2. Description of the Related Art In recent years, with the aging of society, osteoporosis, in which bone density decreases and bone becomes brittle, has become a major problem. When osteoporosis occurs, symptoms such as bending of the bone, severe pain, and fractures are observed. In particular, when the femoral neck is broken, the patient may become bedridden, and an effective treatment method is required.

[0003] As a therapeutic drug for osteoporosis active vitamin D ₃ and the female hormone (estrogen), calcitonin,
Although ipriflavone, bisphosphonate, vitamin K, and the like have been developed, it is often difficult to treat osteoporosis, and it is important to strive for prevention by taking appropriate exercise and paying attention to foods taken daily. To prevent osteoporosis, dietary intake of dairy products and fish that are high in calcium is recommended when it comes to diet. Furthermore, the use of health foods containing calcium, magnesium, vitamin D, and casein phosphopeptide (which is said to have an effect of promoting calcium absorption from the intestinal tract) is also recommended.

[0004] In recent years, the effects of vitamin K on bone have attracted attention, and various uses have been studied. Vitamin K is mainly composed of vitamin K ₁ (phyloquinone) produced by plants and vitamin K ₂ (menaquinone) produced mainly by microorganisms. Vitamin K _{2 also} has menaquinone (MK)-due to differences in the length of side chains. 1-14. Vitamin K is widely present in foods, but its content is minimal. On the other hand, recently, it has been noted that soy isoflavone has a bone strengthening effect. Soy isoflavones include daidzin, glycitin, genistin, daidzein
(daidzein), glycitein, genistein
(genistein), 6 "-0-acetyl daidzin, 6" -0
-Malonyl daidzin, 6 "-0-acetylglycitin,
There are components such as 6 "-0-malonylglycitin, 6" -0-acetylgenistin, 6 "-0-malonylgenistin, etc. Also, succinylated daidzin, succinylated glycitin and succinylated genistin modified by Bacillus natto There is a component of.

[0005]

[Problems to be Solved by the Invention] Vitamin K is contained in a very small amount in foods, but it is contained only in a part of foods such as natto, chlorella, seaweed and green-yellow vegetables in the amount of ppm or more. Absent. However, in each case, the amount is too small to exert an antiosteoporotic effect. Vitamin K is used for the treatment of osteoporosis. However, ingestion of 45 mg per day, which is a therapeutic dose, may rarely cause side effects such as abdominal pain, headache, and rash. With smaller amounts, the therapeutic effect is diminished.

[0006] Soy isoflavone is contained in soybeans and soybean foods in an amount of about 100 to 4,000 ppm, and has been traditionally consumed from soybean foods. However, if an attempt is made to ingest a sufficient amount of soy isoflavone only from a soy food to exert an anti-osteoporosis effect, it is necessary to consume a large amount of soy or soy food, which is a problem in taste. In recent years, products containing high concentrations of soy isoflavones have been developed, but soy isoflavones have a female hormone effect, and there is a concern that ingestion of large amounts may cause enlargement of the uterus or infertility.

[0007] In addition, calcium, magnesium, vitamin D, casein phosphopeptide and the like are used for the purpose of strengthening bones, but their effect of increasing bone strength is low. That is, within the safe intake range,
There is a demand for the development of a functional food having an effect of preventing or improving osteoporosis.

[0008]

Means for Solving the Problems The inventors of the present invention have conducted intensive studies to solve the above-mentioned problems, and as a result, by fortifying both vitamin K and soy isoflavone as active ingredients, more effective and safe prevention of osteoporosis. Alternatively, they have found that a therapeutic composition can be provided and completed the present invention. That is,
The present invention provides vitamin K and soy soflavone as active ingredients.
Containing 100 ppm of vitamin K in dry weight
And the soy isoflavone concentration is 5000 ppm or more
Anti-osteoporosis agent and anti-osteoporosis device characterized by the following
It is a food with the ability. Anti-osteoporosis agent and anti-osteoporosis agent of the present invention
In foods having an osteoporosis function, the mechanism of interaction between both vitamin K and soy isoflavone when acting on bone tissue has not been elucidated.
Probably, the effect of inhibiting the bone resorption of soy isoflavone and the effect of promoting the calcification of vitamin K work in harmony.

[0009]

DETAILED DESCRIPTION OF THE INVENTION Vitamin K used in the present invention
May be either vitamin K ₁ or vitamin K ₂ , and either synthetic or natural products can be used, but when used as food, naturally occurring products are desirable. Also, in the case of vitamin K ₂ derived from a microorganism, vitamin K ₂ derived from Bacillus subtilis typified by Bacillus natto with long eaten it is desirable. In addition, the soy isoflavone used in the present invention is not particularly limited in the production method and composition, and a soybean germ containing isoflavone in a high degree from soybean is taken out and crushed as it is, or purified by an appropriate means. Or daidzin, glycitin, genistin, daidzein, glycitein, genistein, 6 "-0-acetyl daidzin, 6" -0-malonyl daidzin, 6 "-
A product obtained by separating and concentrating 0-acetylglycitin, 6 "-0-malonylglycitin, 6" -0-acetylgenistin, 6 "-0-malonylgenistin, etc. may be used. Natto, miso, soy sauce,
It may be newly produced by performing some modification such as succinylation or hydroxylation after the production of fermented soybean food such as tempe, or may be modified by enzymatic treatment or other methods. When ingesting the composition made according to the present invention, the amount of ingestion varies depending on symptoms, age, and the like, and is not particularly limited.

It is essential that the composition according to the present invention contains both vitamin K and soy isoflavone as active ingredients. In addition to these active ingredients, calcium, magnesium, iron, manganese, Minerals such as copper and zinc, vitamins such as vitamin B group, vitamin D, vitamin E, ubiquinone, and peptides such as isoflavonoids and flavonoids, casein phosphopeptide, and other proteins, lipids, sugars, and oligosaccharides For example, various food ingredients and food additives which are said to be effective for action on bones, and further, chemicals may be contained. In addition, carriers, fillers, colorants, flavors, and the like that are acceptable for pharmaceutical or food production may be used.

[0011] While soy foods Vitamin K ₁ and soy isoflavones are contained trace amounts, for both components exert a synergistic effect, vitamin K concentration is an amount insufficient at all. For example, natto is an excellent food containing the vitamin K ₂ and soy isoflavones, to ingest a sufficient amount of vitamin K ₂ and soy isoflavones to exert effects on bone, it is necessary to eat a large amount This is a matter of taste. Anti-osteoporosis agent and anti-osteoporosis function in the present invention
Is food, vitamin K concentration in dry weight with 100
in ppm or more and it is soy isoflavone concentration 5000ppm or more. Preferably, the vitamin K concentration is 100 ppm or more, and the soy isoflavone concentration is 7000 ppm or more, more preferably, the vitamin K concentration is 200 ppm or more,
And it is more effective if the soy isoflavone concentration is 7000 ppm or more.

[0012]

EXAMPLES Next, the present invention will be described in more detail with reference to examples and product examples, but the present invention is not limited thereto. The following concentrations of soy isoflavone and vitamin K were measured by high performance liquid chromatography.

EXAMPLE The vitamin K having the composition shown in Examples 1 and 2 in Table 1 was used.
A material containing both and soy isoflavone was prepared as follows. Vitamin K ₁ and Vitamin K ₂ used were based on hexane extracts of rapeseed and natto, respectively.
Each was purified to a purity of 98% or more by liquid chromatography and used. The soy isoflavone was used as a raw material from a hot water extract of the soybean hypocotyl and purified by liquid chromatography to a purity of about 91% before use. Its composition is daidzin 22.6%, glycitin 2.5%, genistin 4
6.9%, daidzein 9.8%, glycitein 0.7
%, Genistein 9.1%. Vitamin K ₁ or vitamin K _2, cellulose powder soy isoflavones (Asahi Chemical Industry Co., Ltd.), maltose, alpha-cyclodextrin was added and mixed until uniform in a high-speed mixer, in Examples 1 and 2 Material Was prepared. At the same time, which does not contain both components (Comparative Example 1: control), those containing only soy isoflavone as an active ingredient (Comparative Example 2), those containing only vitamin K ₁ (Comparative Example 3), containing only vitamin K ₂ (Comparative Example 4) was prepared in the same manner as in the example.

[0014]

For each material of the examples and comparative examples,
The following animal experiments were performed to examine the anti-osteoporosis effect. (1) Osteoporosis improvement effect test a. Test Method The ovaries of female SD rats (22 weeks old) were surgically removed to prepare osteoporosis model rats. The ovariectomized rats were divided into 6 groups of 7 rats each, and during the test period of 35 days, every other day (a total of 17 times), Examples 1 and 2 and Comparative Examples 1, 2, 3, and 4 shown in Table 1 were used. 170mg each material
Each of the suspensions was suspended in 1 ml of physiological saline, and orally administered using a magensonde. The feed is CRF, a solid feed for mice, rats and hamsters from Oriental Yeast Co., Ltd.
-1 was used and the feeding and watering method were free intake. During the test period, there was no difference in food intake between each group. After weighing the rats 35 days after the start of the test,
The femur was removed. The femur was used for analysis after removing the adhesive tissue and muscle. After measuring the volume of the femur, it was washed three times with ethanol and then three times with acetone, dried overnight, and then weighed to determine the dry weight of the femur. From the volume and the dry weight, the bone density (dry weight g / volume mm ³ ) was measured. The results are shown in Table 2.

B. Test results The effects of soy isoflavones and vitamin K on ovariectomized rats, which are commonly used as a model for osteoporosis,
The results in Table 2 show that both synergistically improve bone density. Administration of only soy isoflavone (Comparative Example 2) did not improve bone density as compared to the control (Comparative Example 1). On the other hand, even in the administration of only vitamin K ₁ (Comparative Example 3), bone mineral density was not improved. However, when a material in which both soy isoflavones and vitamin K ₁ were fortified as in Example 1 was administered, the bone density was significantly higher than in the control (Comparative Example 1), as well as in Comparative Examples 2 and 3. Improved. That is, by the ingestion of soy isoflavones and vitamin K ₁ at the same time, a synergistic effect is exhibited with respect to the bone, it is clearly visible that the bone density is improved.

Furthermore, in the case of vitamin K _2, Example 2 including both soy isoflavones and vitamin K _2, as shown in Table 2 were observed synergistic effect on bone metabolism. When vitamin K ₂ is administered alone (Comparative Example 4), bone density is significantly improved as compared with the control (Comparative Example 1).
On the other hand, administration of soy isoflavone alone (Comparative Example 2) did not improve bone density. However, when soy isoflavone and vitamin K ₂ were administered simultaneously (Example 2),
Than the effect on bone density of vitamin K ₂ in Comparative Example 4 showed a higher bone density improving effect. In other words, a significantly higher effect was observed when vitamin K ₂ and soy isoflavone were co-present than when they were administered alone. From the above results, according to the present invention, alone is not effective in improving osteoporosis,
Even if the amount of vitamin K is weak, when taken together with soy isoflavone, both components act synergistically,
It was revealed that the anti-osteoporosis effect was significantly promoted.

[0018] ^* p <0.05, ^** p <0.01, ^*** p <0.001: A significant difference is observed from Comparative Example 1 (control). Comparison between Example 1 and Comparative Example 2: A significant difference is observed at a risk rate of p <0.05. Comparison between Example 1 and Comparative Example 3: A significant difference is recognized at a risk rate of p <0.05. Comparison between Example 2 and Comparative Example 4: A significant difference is observed at a risk factor of p <0.001.

Safety Test In order to confirm the safety of the product of the present invention, a mutagenicity test and an acute toxicity test were performed on Examples 1 and 2. (1) Mutagenicity test a. Test method Mutagenicity was examined by a reverse mutation test using bacteria. Test bacteria are Salmonellatyphimurium TA100, TA1535, T
A98, TA1537 and Escherichia coli WP2 uvrA were used. For the test, use Nutrient Broth No.2 (Ox
Add the thawed bacterial solution to an L-shaped test tube containing
What was cultured at 37 ° C. for 10 hours was used as a test bacterial solution. As the test substance, a test substance preparation solution of 50 mg / ml was prepared with water, and used after dilution. As a positive control substance, 2-
(2-furyl) -3- (5-nitro-2-furyl) acrylamide, sodium azide, 9-aminoacridine, 2-aminoanthracene were used. The test consists of an S9mix addition test in which the test substance acts on the test bacteria as it is, and an S9mix addition test for testing the mutagenicity of the metabolite of the test substance produced by the drug metabolizing enzyme (S9mix) possessed by mammals.

B. Test results In the five test bacteria used, no increase in the number of mutant colonies more than twice the solvent control value was observed in neither the S9mix-free test nor the S9mix-added test. , In the test system used,
It was judged to be non-mutagenic (negative).

(2) Acute toxicity test a. Test Method Five female 5-week-old healthy ddy mice were used per group. After fasting for 18 hours, the test substance was orally administered by gavage using a magensonde so that the test substance became 5 g / kg with respect to the body weight of the individual. While measuring the body weight of each individual with the administration day as 0 day, the general symptoms and the state of life and death for 7 days were observed. Water and food were provided ad libitum during this time. At the same time, the condition of the uterus was observed.

B. Test Results In this test, no deaths were observed in the mice, and the weight increased steadily. The appearance was not different from a normal mouse. Therefore, the LD ₅₀ of both test substances was 5
g / kg and proved to be a safe composition. After dissection 7 days later, the condition of the uterus was observed, and no abnormality was observed.

From the results of the performance test and the safety test, the anti-osteoporosis composition formulated according to the present invention can simultaneously take safe amounts of vitamin K and soy isoflavone as foods. It has been found that an excellent bone strengthening action that cannot be obtained can be obtained.

[0024]

[Product example]

Product Example 1 Capsule Vitamin K ₂ purified from butter is dissolved in edible oil,
To produce a vitamin K _2-containing oil. The soybean hypocotyl was crushed to obtain a crushed soybean hypocotyl. In addition, vitamin E is provided for the purpose of protecting the oxidation of oil in the product and fortifying at the same time,
A stock solution of a capsule was prepared based on the following formulation. [Formulation] (1) vitamin K ₂ contained oil (vitamin K concentration 300 ppm) 130 mg (2) soybean hypocotyl pulverized product (soy isoflavone concentration 28,500ppm) 50mg (3) Vitamin E (alpha-tocopherol concentration 98%) 20 mg (dry weight (Vitamin K concentration: 195 ppm, soy isoflavone 7,125 ppm) The mixture per capsule was mixed well so that the above ratio was obtained, and the capsules were encapsulated with gelatin capsules. The obtained capsule was highly stable, excellent in storage stability and portability.

Product Example 2 Drink Vitamin K ₂ was purified from natto by liquid chromatography. From the soybean hypocotyl, a fraction containing soy isoflavone was extracted with ethanol and purified to a concentration of 91% by preparative liquid chromatography. [Prescription] Ingredients Ingredients (1) Vitamin K ₂ (concentration: 90%) 0.03 g (2) Soy isoflavone (concentration: 91%) 1.5 g (3) Dextrin powder 20 g (4) Milk 1000 g (per dry weight) (Vitamin K concentration: 189 ppm, soy isoflavone concentration: 9,545 ppm) The above raw materials were mixed with a mixer to obtain a uniform milk drink.
The resulting beverage has good taste and can be taken continuously.

Product Example 3 Tablets Using rapeseed as a raw material, high performance liquid chromatography
Vitamin K ₁ was purified. As the soy isoflavone, a soybean hypocotyl hot water extract roughly purified by column chromatography was used. In addition, the following auxiliaries and materials for health food were purchased and used. [Formulation] material amount of vitamin K ₁ 1 mg Soybean isoflavone (concentration 70%) 30 mg scallop powder 70mg lactose 120mg reduced maltose 15mg Corn starch 12mg cellulose powder 15mg dry yeast powder 10mg vitamin D ₃ 18 IU (International Units) (dry weight per (Vitamin K concentration: 3,636 ppm, soy isoflavone concentration: 76,364 ppm) The raw materials were mixed well in a mixing ratio of not less than the above, and the tablets were obtained by tableting. The obtained tablets can obtain calcium from scallop powder as well as soy isoflavone and vitamin K, and can be expected to have an excellent preventive effect against osteoporosis as a health food.

[0027]

The anti-osteoporosis composition formulated according to the present invention can obtain a more effective anti-osteoporosis effect by enhancing both vitamin K and soy isoflavone.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification code FI A23L 2/38 A23L 2/38 P 2/52 A61K 31/12 A61K 31/12 31/35 31/35 35/78 35/78 A61P 19/10 A61P 19/10 A23L 2/00 F

Claims (4)

(57) [Claims] 1. A composition comprising vitamin K and soy soflavone as active ingredients, wherein the concentration of vitamin K in the dry weight is 100%.
ppm or more and soy isoflavone concentration 5000 ppm or more
An anti-osteoporosis agent, characterized in that: 2. The anti-osteoporosis agent according to claim 1, wherein the vitamin K is natural vitamin K. 3. Vitamin K is derived from Bacillus subtilis.
_2. The anti-osteoporosis agent according to claim 1, which is ₂ . 4. Vitamin K and soybean sofra as active ingredients
And the concentration of vitamin K in the dry weight is 100
A food having an anti-osteoporosis function, characterized in that the concentration of soy isoflavone is 5,000 ppm or more.