KR20050003989A - Egg derivatives having bone-strengthening effects - Google Patents

Abstract

PURPOSE: An egg-derived bone-strengthening composition containing an egg-derived water-soluble fraction having the activity of promoting the proliferation of osteoblasts, promoting calcification, enhancing chondrocyte proliferation and promoting bone growth is provided. It can be taken orally, added to food such as milk products, milk drinks, drink yogurt, coffee drink, fruit drink, jelly, pudding, cookie, biscuit and wafer having bone-strengthening action and is useful as raw material for food, drug or feed for preventing and improving osteoporosis. CONSTITUTION: The bone-strengthening composition contains an egg-derived water-soluble fraction as an effective component. The egg-derived water-soluble fraction is obtained by defatting treatment of a whole egg or egg yolk powder with an organic solvent and then extracting in warm water or 0.5% sodium chloride, or adding an aqueous carrageenan solution to a whole egg or egg yolk liquid.

Description

Egg derivatives having bone-strengthening effects

The present invention relates to a composition containing an egg-derived water-soluble fraction having bone growth and bone strengthening action. The present invention also relates to a food or beverage, a medicine, or a feed containing the egg-derived water-soluble composition. Since the oocyte-derived water-soluble fraction of the present invention has the effect of proliferating osteoblasts and promoting the proliferation of chondrocytes in the bone growth plate, it is possible to prevent or treat bone diseases caused by the breakdown of bone metabolism, and also to young people such as elementary school students. It is useful for strengthening and growing bone during growth.

Recently, as osteoporosis increases, the increase of various bone diseases such as an increase in fracture rate of young people is emerging as a social problem. It is estimated that there are about 10 million osteoporosis patients nationwide, and as the aging progresses, the number of patients is expected to increase to 16 million in 2010. Osteoporosis is a disease in which bone formation and bone destruction are out of balance due to the reduction of the female hormone (Estrogen hormone), mainly in postmenopausal women, but even men are seeing a decrease in bone mass with aging. There are about 2 million osteoporosis patients in this age group. In addition, the fracture rate of elementary and middle school students has more than doubled in recent 20 years. In terms of preventing osteoporosis, it is important to increase bone mass and to obtain high maximum bone mass from a young age. As such, maintaining bone health is a global concern regardless of gender or age.

Background Art In the past, as a method of preventing or treating bone diseases, calcium diffusion by diet, light exercise, sunbathing and drug treatment have been performed. Although calcium salts such as calcium carbonate and calcium phosphate, natural calcium preparations such as right bone meal, eggshell and fish bone meal are used for calcium diffusion by diet, solubility, water absorption, In terms of taste, it is not suitable for oral ingestion. Proper exercise increases bone mass and strengthens bones, so walking and walking make bones healthy, but when your body is weak, light exercise can be cumbersome, and hardly even older people can hardly exercise. Sunbathing is good in terms of the spread of active vitamin D3, but this is not enough. Bisphosphonate or calcitonin is used for drug administration and is known to be effective in the treatment of osteoporosis. However, these substances are specialty medicines, and they require special attention, and there is a risk of side effects such as tinnitus, headache, and anorexia, so they are not usable as a continuous food ingredient.

It is an object of the present invention to provide a food material that is a bone strengthening agent that can be continuously ingested on a daily basis, which has a bone strengthening action, which is safe and inexpensive, and which can be widely used in food.

Eggs contain all the materials that form a living body to be able to hatch into chicks when kept at 37 ° C. for three weeks. It is a biocapsule that produces life. In addition, eggs have been used as food since ancient times, and their nutritional value is high and safe, so the use value as a food material is quite high.

Accordingly, the present inventors have searched for the bone growth and bone mineralization promoting components for various fractions of eggs, and for the first time found that the yolk (water yolk) water-soluble fraction has osteoblast proliferation and calcification promoting activity, Enhancement of bone growth and improvement of bone metabolism by ingestion have been found to complete the present invention.

That is, the bone-enhancing composition of the invention described in claim 1 (hereinafter referred to as "invention 1") contains, as an active ingredient, an aqueous fraction derived from eggs having osteoblast proliferation promoting activity, bone growth, and osteoporosis. It is characterized by one.

In the present invention 2, the egg yolk soluble fraction is egg yolk degreased by at least one organic solvent in ethyl alcohol, isopropanol, nucleic acid, or other organic solvents used in food processing. It is preferably a water soluble fraction extracted from.

Moreover, in this invention 3, it is preferable that the said egg yolk soluble fraction is an extract obtained by adding carrageenan liquid to whole egg or egg yolk liquid.

In addition, it is preferable that the food-drinks, medicine, or feed of this invention 4 mix | blended and used the egg yolk (yield) water-soluble composition obtained by this invention 1, invention 2, and invention 3.

According to the present invention, it is possible to provide a safe, inexpensive and routinely ingestible bone strengthening material and food, medicine, or feed.

1 is a graph comparing the growth rate of the tibia with respect to each feed intake group when the rats were fed the rats (Rat), which is usually added to the feed and the yolk soluble fraction.

Figure 2 is a diagram showing the appearance of BrdU intercalated into the cartilage cells of the tibial growth plate in each breeding group administered the feed, egg yolk soluble fraction feed. The BrdU labeling rate of the sample intake group was significantly increased compared to the control group. In the normal feed intake group shown in the figure, the nuclei of the three cells indicated are stained, and the yolk soluble fraction added feed intake group is stained in 11 cells.

Eggs used in the present invention can be used for all eggs, such as chicken, duck, quail, eggs were used in terms of productivity.

Egg eggs, egg yolk, egg white, egg powder, egg yolk powder, egg white powder (卵 白 粉末), degreasing egg powder (脫脂 全 卵 粉末) and degreasing egg yolk powder (脫脂 卵 黃 粉末) can be used, of which degreasing egg powder or degreasing egg yolk powder was used.

The bone-reinforced composition of the present invention can be obtained by using an organic solvent which can be used for food processing from whole egg powder, degreasing and extracting the powder with hot water or 0.5% sodium chloride aqueous solution and then obtaining it from the supernatant. The organic solvent used for degreasing can be used as ethyl alcohol, isopropanol, hexane, and other organic solvents that can be used for food processing. However, in terms of simplicity and safety, Is preferred.

In addition, the bone strengthening composition can be obtained as an extract obtained by adding a natural rubbery aqueous solution such as gum arabic, carrageenan, xanthan gum, or the like to a whole egg yolk solution or egg yolk solution. The natural gum-based aqueous solution used here can be used either gum gum, carrageenan, xanthan gum, etc. Particularly preferred.

The bone strengthening composition of the present invention is applied to the prevention or improvement of osteoporosis and various bone diseases, and its effective intake is 100 g to 6 g in terms of egg yolk water soluble fraction powder per adult. In addition, the bone-reinforced composition of the present invention was extracted from food and did not show acute toxicity in experiments by rats.

The osteoporosis composition of the present invention may be treated with proteolytic enzymes as necessary. The proteolytic enzyme is not particularly limited as long as it is a proteolytic enzyme that can be used in food, and any acidic protease, neutral protease, or alkaline protease can be used. In the present invention, neutral protease is preferably used. Examples of the neutral protease include orientase (trade name, manufactured by HBI Corporation), flavazyme (trade name, manufactured by Novonol Disk), and the like.

The osteoporosis composition recovered in this way is usually powdered and used. The powdering is powdered by a lyophilization or spray dry (spray drying) method and usually orally administered as food.

In addition to the components described above, the osteoporosis composition of the present invention is a component used in conventional foods such as excipients, emulsifiers, stabilizers, sweeteners, pH adjusters, thickeners, proteins, peptides, amino acids, lipids, polysaccharides, oligosaccharides, etc. It may include.

The bone-reinforcement composition of the present invention is not only put into food and drink, but also produced in various forms depending on the dosage form. Egg is a safe food material that has been used as a food for a long time, and it is a natural material without side effects even if it is used continuously, and can be used as a medicine or a feed.

Hereinafter, the present invention will be described in more detail with reference to Examples.

Example 1 Extraction with Organic Solvents

2.5 liters of ethyl alcohol (ethanol) was added to 500 g of egg yolk powder, and the mixture was stirred for 30 minutes to recover a solid. This operation was repeated three times to perform degreasing from egg yolk. It was air dried to obtain 284 g of skim egg yolk (yolk). 3 L of warm water at 37 ° C. was added to the degreasing egg yolk powder, mixed with a stirrer while maintaining the temperature, and centrifuged at 8000 rpm for 30 minutes to obtain a clear liquid at the top. It was freeze-dried to obtain 13.4 g of yolk (yellow) water-soluble composition.

Example 2 Extraction by Carrageenan

3L of 1% carrageenan solution was added to 500 g of egg yolk powder, stirred gently, and left to stand for 30 minutes to show aggregation. This was centrifuged at 8000 rpm for 30 minutes to obtain a clear liquid at the top. This was freeze-dried to obtain 14.8 g of yolk (yellow) water-soluble composition.

The molecular weight of the water-soluble composition obtained in Examples 1 and 2 was analyzed by gel filtration chromatography under the following conditions.

Column: YMC-Pack

Diol 200 (6.0 × 300 mm) (brand name, YMC Co., Ltd.)

Eluent: 0.2 M potassium phosphate buffer (pH 6.9)

Flow rate: 0.7ml / min

Detection wavelength: 280nm

As a result, all of the yolk (water yolk) water-soluble compositions of Examples 1 and 2 showed the distribution of major peaks of the same aspect with a molecular weight of 10,000 to 200,000.

Test Example 1 (Measurement of Osteoblast Proliferation Activity)

Osteoblast proliferation activity of the yolk (water yolk) water-soluble composition obtained in Example 1 was measured as follows. InM1.2 cells of human osteoblasts were cultured at 37 ° C. and 5% CO ₂ -95% air in an α-MEM medium containing 10% FBS, and cultured until confluent. Cells were collected by treatment with trypsin, and the concentration was adjusted to 1 × 10 ⁴ / mL using the culture solution. 0.5 mL of this cell preparation was dispensed in a 24-well plate and incubated under 37 ° C and 5% CO ₂ -95% air. The next day, the sample was added to each well so that it became 100, 25, and 12.5 ppm, and 2 mmol / L of alpha-glycerol phosphate which is a calcification promoter was added. The sample was dissolved in 50% DMSO and the solvent added to 1% of the culture. The cultures were exchanged daily and after 20 days the number of osteoblasts was measured using MTT. The proliferation promoting activity of osteoblasts was expressed as a relative value when the proliferation number of the control group was 100. The results are shown in Table 1.

TABLE 1

Test Example 2 (Bone Growth Promoting Activity)

The bone growth promoting activity of the yolk (water yolk) water-soluble composition obtained in Example 1 was measured as follows. Bone growth promoting activity was assessed using Sprague-Dawley rats. Samples were dissolved in distilled water at a concentration of 20 mg / ml and rats were ingested at 20 and 100 mg / kg body weight / day. Rat water and feed were freely ingested during the test period, and MF (trade name, oriental yeast company) was used as a basic feed. Tetracycline, a fluorescent substance, was administered on days 3 and 5 of the sample intake period (1 week), respectively. In this test example, tetracycline was chelate-bonded with calcium and used to deposit on bone, and fluorescence lines formed on bone were observed to determine the tibial egg protein composition. The growth promoting effect was measured. The tibia was removed, and the length between each fluorescent line formed at the lower part of the tibial growth plate and the bone angiogenesis, ie, the grown length, was measured by administration of tetracycline. Bone elongation was compared with the case where the yolk (yellow yolk) -derived water-soluble composition was given and the base sample (control) were evaluated, and the promoting effect was evaluated. As a result of the microscopic observation, it was confirmed that the egg yolk-derived water-soluble composition group had a 30% wider spacing between the two layers of the dye band for the control group, thereby promoting bone resorption activity (FIG. 1). .

Test Example 3 (Animal Test Evaluating Cartilage Cell Proliferation)

Cartilage cell proliferation in vivo in the yolk (water yolk) water-soluble composition obtained in Examples 1 and 2 was performed as follows. Evaluation of cartilage cell proliferation was carried out using a bromo deoxyuridine (BrdU) which measures the DNA synthesis cycle, that is, cell proliferation in the cell cycle. The egg yolk (yolk) water-soluble composition obtained in Examples 1 and 2 was ingested into Sprague-Dawley rats at a rate of 100 mg / kg body weight / day. During the test period, rats were freely given water and feed, and MF (trade name, Oriental Yeast Co., Ltd.) was used as feed. After ingestion period (1 week), BrdU was intraperitoneally administered (Brdu 50mg / kg), and tibias were extracted from rats anesthetized 2 hours after administration, and BrdU influx into the chondrocytes of growth plate was obtained using anti-BrdU antibody. It measured by the immunostaining method. In the sample intake group, BrdU inflow rate activity was significantly higher than the control. This indicates that the proliferation activity of chondrocytes is increased by sample ingestion, and that bone growth is promoted (FIG. 2).

Example 4 (soft drinks)

A soft drink containing an egg yolk (yolk) water-soluble composition prepared in Example 2 was prepared. That is, the composition is mixed 15.0% isomerized sugar, 10% fruit juice, 2.0% egg yolk (water yolk) water-soluble composition, 0.1% flavor, 0.1% calcium (calcium), 72.8% water, using a plate sterilizer , Sterilized for 15 seconds at 90 ℃ to prepare a soft drink having a bone strengthening action.

Example 5 (Yogurt)

Yogurt containing the yolk (water yolk) water-soluble composition adjusted in Example 2 was adjusted. That is, a yogurt having a bone strengthening effect was prepared by mixing raw materials having a composition of yolk (yolk) water-soluble composition 3.0%, sucrose 7%, flavor 0.1%, and yogurt 89.9%, and filling the container.

Example 6 Cheese

Process cheese containing the yolk (yellow) water-soluble composition adjusted in Example 2 was adjusted. That is, Gouda cheese 35%, cheddar cheese 35%, Parmesan cheese 20%, egg yolk aqueous composition 2.0%, calcium phosphate 1.0%, water 7.0% Each raw material was mixed to include, and then emulsified at an emulsification temperature of 85 ° C. to produce a process cheese having a bone strengthening action.

Example 7 (Capsules)

Each raw material was blended to contain 60% of the yolk (water yolk) water-soluble composition obtained in Example 2, 30% cornstarch, and 10% lactose, and filled into gelatin capsules (200 mg per capsule). Thus, a capsule having a bone strengthening action was prepared.

Example 8 (Tablet)

After blending each raw material so as to contain 60% of the yolk (water yolk) water-soluble composition obtained in Example 2, 18% of reduced malt sugar, 18% of crystalline cellulose, and 4% of sucrose ester, tableting (300 mg per tablet) To prepare a tablet having a bone strengthening action.

As described above, the egg-derived water-soluble composition of the present invention has osteoblast proliferation promoting and calcification promoting activities. In addition, it has the activity of promoting the proliferation of chondrocytes and the growth of bone. In addition, the bone enhancer of the present invention can be taken orally, dairy products such as cheese (cheese), butter (butter), fermented milk, milk beverage, drink yogurt (coffee), beverages such as juice, Food and drinks that have a bone strengthening effect by being added to various types of food and beverages such as jelly, pudding, cookies, biscuits, wafers, and frozen foods. Can be made with

It can also be used as a supplement for capsules, tablets and the like, and is useful as a material for food and drink, medicine or feed for the purpose of preventing osteoporosis and improving symptoms.

Claims (4)

An osteoporosis composition containing an oocyte-derived water-soluble fraction having osteoblast proliferation, bone growth, and bone mineralization as an active ingredient. The egg-derived water-soluble fraction of claim 1, wherein the whole egg or egg yolk powder is degreased with an organic solvent and then extracted with hot water or 0.5% sodium chloride aqueous solution. Osteoporosis composition of Claim. The bone strengthening composition according to claim 1, wherein the egg-derived water-soluble fraction according to claim 1 is obtained by adding a carrageenan aqueous solution to whole egg or egg yolk solution. A food or beverage, a medicine, or a feed comprising the osteointestinal composition according to claim 1, 2 or 3.