JPH10298082A - Osteo enhancer - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an osteo enhancer having osteo enchancing effect, capable of inhibiting and treating several diseases such as bone fracture, rheumatism, articular inflammation, lumbago, osteoporosis, etc., by containing deoxyribonucleic acid as an active ingredient. SOLUTION: This osteo enhancer uses deoxyribonucleic acid(DNA) obtained from soft roe, etc., of salmon, trout, cod, etc., as an active ingredient. The molecular weight of the DNA is preferably above 1000. The osteo enhancer using DNA as the active ingredient, can be formulated to afford an osteo enhancing foods, eatables, drinkables and feedstuff for animals. The content of the DNA is 0.1-10% in the case of medicines. The DNA is administered at a daily does as 0.5-1.5 g for adults as medicines.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a bone strengthening agent containing deoxyribonucleic acid as an active ingredient. Also, the present invention
The present invention relates to a medicine, a food and drink, and a feed, which are provided with a bone strengthening action by blending a bone strengthening agent containing deoxyribonucleic acid as an active ingredient.
Since the bone strengthening agent of the present invention has a bone strengthening effect, fractures,
It is useful for prevention or treatment of various bone diseases such as rheumatism, arthritis, back pain, and osteoporosis.

[0002]

2. Description of the Related Art In recent years, with the aging of society, the number of people suffering from various bone diseases such as fractures, rheumatism, arthritis, low back pain, and osteoporosis is rapidly increasing, and the prevention and treatment methods thereof are rapidly established. Have been. Various bone diseases such as fractures, rheumatism, arthritis, low back pain, and osteoporosis are considered to be caused by insufficient calcium intake, reduced calcium absorption capacity, and hormonal imbalance after menopause. Therefore, in order to prevent various bone diseases accompanying aging, it is extremely important to obtain as much bone mass as possible during childhood and adolescence and to increase the maximum bone mass. And
In order to increase bone mass, it is important to consume more calcium. However, in order to prevent and treat various bone diseases, it is considered desirable not only to take calcium but also to take vitamin D and other nutrients in a well-balanced manner. Recently, it has been revealed that vitamin K known as a blood coagulation factor also plays an important role in bone metabolism, and vitamin K has been used as a therapeutic agent for osteoporosis. Thus, it is clear that various substances are involved in bone and calcium metabolism.

[0003] On the other hand, deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) are known to be contained in cells of all living organisms and are high molecular substances that control genetic information. Nucleotides that are constituents of these nucleic acids are not only constituents, but also ATP, which is a common energy source for all living organisms, or cAMP and cGMP, which are metabolic regulators, and are also coenzymes. N
It plays an important role in vivo as AD and FAD.

[0004] Thus, despite the fact that nucleic acids, which are essential substances of life, are ingested daily from foods such as cereals, vegetables, meats, fish, etc., the nutritional value of nucleic acids has been mostly watched until recently. On the contrary, it was not always evaluated as a cause of gout due to overdose. However, in the 1980s, there were many reports on the biological usefulness of ingesting nucleic acids and their components, and the idea that nucleic acids are the sixth essential nutrient has begun to appear.

[0005] When a nucleic acid is ingested, it is decomposed and absorbed into nucleotides and nucleosides in the stomach and intestine. In an experiment of oral administration of nucleic acid using mice, 70% of nucleotides and nucleosides were absorbed one hour after administration, of which 15%
Is taken up in the tissue, and the distribution in the tissue 4 hours after administration is 47% in the gastrointestinal tract and in the skeleton such as head, skin, muscle and bone.
There is a report that it is 40%. As for the function of this nucleic acid in the gastrointestinal tract, animal experiments using rats have confirmed that the intestinal villi can be lengthened, the protein and DNA content can be increased, and the intestinal mucosal growth can be activated to prevent mucosal atrophy. It is known that nucleic acids are deeply involved in intestinal growth and maturation. It is also known that ingestion of nucleic acids causes intestinal flora such as bifidobacteria which plays an important role in the intestine to grow. Thus, it has become clear that ingestion of nucleic acids causes morphological changes in the intestine. In addition, yeast-derived RN
A nutritional composition for promoting absorption of calcium containing A as an active ingredient has also been proposed (Japanese Patent Publication No. 7-100657).
.

[0006]

SUMMARY OF THE INVENTION We have found that fractures,
In order to find a substance that has a bone-strengthening action useful for the prevention or treatment of various bone diseases such as rheumatism, arthritis, back pain, and osteoporosis, we have been conducting intensive research and found that DNA obtained from milt and the like has a bone-strengthening action. Was found. And
It has been found that by adding this DNA to medicines, foods and drinks and feeds, a bone strengthening action can be imparted, and the present invention has been completed. Therefore, an object of the present invention is to provide a bone strengthening agent containing DNA as an active ingredient. Another object of the present invention is to provide a medicine, a food and drink, and a feed that have a bone-strengthening effect by blending a bone-strengthening agent containing DNA as an active ingredient.

[0007]

According to the present invention, DNA is used as an active ingredient of a bone enhancer. This DNA can be prepared from milt or the like according to a general nucleic acid extraction method such as salt extraction, organic solvent extraction, and surfactant extraction. In addition, a DNA-rich fraction can be obtained by removing the milt of the milt, treating the homogenate prepared by crushing with a homogenizer with a proteolytic enzyme to degrade the protein, and removing the precipitate. If the molecular weight of DNA is too small, DN
Since A may be decomposed into sugars, bases, and phosphates and its effectiveness may be reduced, it is preferable that the molecular weight of DNA is 1,000 or more. Examples of the proteolytic enzymes used to decompose the milt homogenate protein include enzymes derived from animals and plants, such as trypsin and papain, or Aspergillus genus and Bacillus ( Bacillus).
Examples include enzymes derived from microorganisms such as genus illus and Rhizopus .

There is no particular limitation on milt used as a raw material in preparing DNA, and salmon, trout, cod,
It is preferable to use those collected from fish and shellfish such as herring, squid and scallops, but it is particularly preferable to use milt of fishes with high catches such as salmon, trout and cod from the viewpoint of effective use of resources. It can be said that.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention is a bone strengthening agent containing DNA as an active ingredient. With respect to this bone enhancer, the DNA may be used as it is as a bone enhancer, or a formulation component usually used for formulation, for example, a bulking agent, a diluent, an excipient such as a solvent or a filler, Solubilizers, solubilizers, emulsifiers, suspending agents, dispersants, binders, lubricants, auxiliaries such as coating agents and sustained release agents, or antioxidants, preservatives, brighteners, sweeteners Agents, coloring agents, and additives such as flavoring agents, and mixed with tablets, capsules, granules, pills,
Various dosage forms such as drinks, syrups, and liquids may be used as medicaments imparting bone-strengthening action.

[0010] The present invention also provides a bone strengthening agent containing DNA as an active ingredient, in the form of dairy foods such as cheese, butter and fermented milk, drinking milk, drink yogurt, coffee drinks, fruit juices and other drinks, jelly, pudding and cookies. , Biscuits, confectioneries such as wafers, and frozen foods and other various foods and drinks, which may be used as foods and drinks having a bone strengthening effect. Furthermore, it can be blended with animal feed to give a feed that has a bone strengthening effect.

When the bone enhancer of the present invention is blended in medicines, foods and drinks and feeds, the blending amount is not particularly limited, but it is preferable to blend it in such a manner that the DNA content is 0.1 to 10%. I can say. In the case of preventing or treating various bone diseases such as fractures, rheumatism, arthritis, back pain, osteoporosis, etc., in order to exert a bone strengthening effect, in the case of an adult, it is necessary to take about 0.5 to 1.5 g of DNA per day for adults. Is preferred.

Next, the present invention will be described in more detail with reference to Examples and Test Examples.

Example 1 1 kg of salmon milt (containing 12% of nucleic acid) from which the rind was removed was washed with water, dehydrated, and 3 L of water was added thereto to prepare a homogenate with a homogenizer. A 0.05N sodium hydroxide solution was added to the homogenate to adjust the pH to 6.
After adding 20 g of papain and stirring at 35 ° C. for 2 hours to decompose the protein, the mixture was kept at 80 ° C. for 30 minutes to inactivate the enzyme and terminate the decomposition. After the digestion, the precipitate was removed by centrifugation.
% Ethanol was added to precipitate the DNA. Then, after the aqueous ethanol is removed by filtration, it is freeze-dried, and D
130 g of a bone enhancer composed of NA powder was obtained. Note that 1 g of this bone enhancer contained 850 mg of DNA.

[0013]

Example 2 1 kg of salmon milt (containing 12% of nucleic acid) from which the rind was removed was washed with water, dehydrated, and 3 L of water was added thereto to prepare a homogenate with a homogenizer. A 0.05N sodium hydroxide solution was added to the homogenate to adjust the pH to 6.
After adding 20 g of papain and stirring at 35 ° C. for 2 hours to decompose the protein, the mixture was kept at 80 ° C. for 30 minutes to inactivate the enzyme and terminate the decomposition. After the digestion, remove the precipitate by centrifugation, add an equal amount of neutral phenol to the resulting supernatant, gently shake for 20 minutes at room temperature, remove the precipitate by centrifugation, and collect the supernatant did. An equal volume of a neutral phenol: chloroform = 1: 1 solvent was added to the supernatant, and the mixture was gently shaken at room temperature for 20 minutes. The precipitate was removed by centrifugation, and the supernatant was recovered. An equal volume of chloroform was added to the supernatant, and the mixture was gently shaken at room temperature for 20 minutes. Then, the precipitate was removed by centrifugation, and the supernatant was recovered. The DNA was precipitated by adding 2.5 volumes of 95% ethanol to the supernatant. Then, the aqueous ethanol was removed by filtration, and the precipitate was washed with 70% ethanol. Then, the ethanol was removed by filtration, freeze-dried, and
115 g of a bone enhancer composed of A powder was obtained. In addition, 1 g of the bone enhancer contained 980 mg of DNA.

[0014]

Test Example 1 1.02% of the bone strengthening agent obtained in Example 2, casein 20%, corn starch 15%, cellulose 5%, corn oil 5%, sucrose 49.18%, DL-methionine 0.3%, mineral mixture 3.5% and An experimental feed was prepared at a mixing ratio of 1% of a vitamin mixture, and was subjected to an animal experiment to confirm the bone strengthening action of DNA.

A 7-week-old male SD rat was used as an experimental animal, and was bred for 1 week after preliminarily bred for 1 week and then fed with a feed containing the bone strengthening agent of the present invention for 4 weeks. Four weeks after the start of the administration of the experimental feed, the femur and the fourth lumbar vertebra were removed, and the bone fracture stress of the femur and the bone density of the fourth lumbar vertebra were measured. The bone fracture stress of the femur was measured by a three-point bending method using a bone fracture strength measuring device (Rheometer Max; manufactured by I-Techno Co., Ltd.).
The bone density was measured by a DXA apparatus (DCS-600A; manufactured by Aloka) by a method using two wavelengths of X-rays (DXA method). In addition, as a comparative example, a similar animal experiment was performed on a feed not containing the bone enhancer of the present invention and a feed containing RNA instead of the bone enhancer of the present invention. FIG. 1 shows the bone breaking stress of the femur and FIG. 2 shows the bone density of the fourth lumbar vertebra.

According to this, the group to which the feed containing the bone enhancer of the present invention was administered was the group to which the feed not containing the bone enhancer of the present invention was administered, and R was replaced by the bone enhancer of the present invention.
It became clear that the bone breaking stress was higher and the bone density was higher than the group to which the diet containing NA was administered.

[0017]

Example 3 The bone strengthening agent of the present invention obtained in Example 1 was added so that the amount added per 100 mL of raw milk was 1 g, and 120 kg
After homogenizing at a pressure of / cm ² , the milk was sterilized at 120 ° C. for 4 seconds to produce milk having a bone strengthening effect. In addition, 0.8 g of DNA was contained in 100 g of the milk provided with the bone strengthening action.

[0018]

Example 4 Skim milk powder was dissolved in water to a solid content of 12%, sterilized by heating at 90 ° C. for 20 minutes, cooled to 25 ° C., and cooled to 25 ° C. to obtain Lactobacillus acidop.
hilus) and Streptococcus thermophilus (Strep)
tococcus thermophilus ) and the lactic acidity is 1.0
%, And when the pH reached 4.3, the mixture was cooled to 5 ° C. to prepare a starter culture. Then, this starter culture was inoculated into sterilized raw milk (fat content: 3.5%) by 5%, and 1.5 g of the bone strengthening agent of the present invention obtained in Example 1 was added thereto. To give yogurt. In addition, yogurt 10 which gave this bone strengthening action
0 g contained 1.1 g of DNA.

[0019]

Example 5 A dough having the composition shown in Table 1 was prepared according to a conventional method, molded, and then roasted to produce a biscuit having a bone strengthening effect.

[0020]

[Table 1] 小麦 Flour 50.0 (% by weight) Sugar 20.0 Salt 0.5 Margarine 12.5 Egg 12.5 Water 2.5 Obtained in Example 1 Bone enhancer 2.0 ─────────────────────────

Incidentally, 1.5 g of DNA was contained in 100 g of the biscuit provided with the bone strengthening action.

[0022]

Example 6 Tablets of a bone enhancer having the composition shown in Table 2 were produced according to a conventional method.

[0023]

[Table 2] 結晶 Hydrous glucose dextrose 88.5 (% by weight) Bone enhancer obtained in Example 2 10.0 Sugar ester 1.0 Fragrance 0.5 ─────────────────────────

Incidentally, DN was contained in 100 g of the tablet of this bone strengthening agent.
A9.7g was contained.

[0025]

Example 7 According to a conventional method, a dog breeding feed (dog food) having a bone-strengthening effect having the composition shown in Table 3 was produced.

[0026]

[Table 3] ───────────────────────── Soybean meal 11 (wt%) skim milk powder 14 soybean oil 4 corn oil 2 palm oil 2 corn starch 30 Flour 15 Bran 8 Vitamin mixture 2 Mineral mixture 9 Cellulose 2 Bone enhancer obtained in Example 2 1

It should be noted that 0.9 g of DNA was contained in 100 g of the dog food provided with the bone strengthening action.

[0028]

EFFECTS OF THE INVENTION Since the bone augmenting agent of the present invention has a bone augmenting action, it is useful for the prevention or treatment of various bone diseases such as fracture, rheumatism, arthritis, back pain, osteoporosis and the like, and is useful in medicine, food and drink or feed. By blending, a bone strengthening action can be imparted.

[Brief description of the drawings]

FIG. 1 shows the measurement results of the bone fracture stress of the femur in the animal experiment of Test Example 1.

FIG. 2 shows the measurement results of the bone density of the fourth lumbar vertebra in the animal experiment of Test Example 1.

Claims (4)

[Claims] 1. A bone strengthening agent comprising deoxyribonucleic acid as an active ingredient. 2. The bone strengthening agent according to claim 1, wherein the deoxyribonucleic acid is a milt-derived deoxyribonucleic acid. 3. The bone enhancer according to claim 1, wherein the deoxyribonucleic acid is a deoxyribonucleic acid having a molecular weight of 1,000 or more. 4. A medicament, a food or drink, and a feed, which has a bone-strengthening effect by blending the bone-strengthening agent according to any one of claims 1 to 3.