JP2001346519A - Method for producing fraction containing high content of milk basic cystatin and decomposed product thereof - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a fraction containing high content of milk basic cystatin and a decomposed product of the fraction containing high content of the milk basic cystatin for prophylaxis and amelioration of various kinds of bone diseases such as osteoporosis, an osteoarthropathy such as rhematism, and periodontal disease, capable of being used as a food and drink, a medicine and a feed. SOLUTION: This fraction containing the high content of the milk basic cystatin is produced by bringing a basic protein composition derived from a milk into contact with an anion exchange resin and collecting a faction not absorbing to the resin and then bringing the fraction into contact with a cation exchange resin and eluting a fraction absorbing to the cation exchange resin with an eluent and the decomposed product of the fraction containing the high content of the milk basic cystatin is produced by decomposing the obtained fraction containing the high content of the milk basic cystatin with a protease.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a method for producing a milk-basic cystatin-rich fraction from a milk-derived basic protein composition using an ion exchange resin. The present invention also relates to a method for producing a milk-basic cystatin-rich fraction hydrolyzate by decomposing a milk-basic cystatin-rich fraction obtained by the above method with a protease. Furthermore, the present invention relates to various bones such as osteoporosis and the like containing the milk-basic cystatin-rich fraction and / or the milk-basic cystatin-rich fraction decomposed product and, if necessary, calcium and / or vitamins. The present invention relates to a food and drink, a medicine or a feed for preventing and improving diseases and osteoarticular diseases such as rheumatism and periodontal diseases.

[0002]

2. Description of the Related Art In recent years, various bone diseases such as osteoporosis, bone fractures and back pain have been increasing with aging. In bone tissue, bone formation and bone resorption are constantly performed, and at a young age, the balance between bone formation and bone resorption is balanced. However, as the age increases, the balance tends to bone resorption due to various causes. If this continues for a long period of time, the bone tissue becomes brittle, resulting in various bone diseases such as osteoporosis, fracture and back pain. It is considered that various types of bone diseases can be prevented if uncoupling that tends to resorb bone can be prevented. Conventionally, as methods for preventing and treating various bone diseases, 1) supplementation of calcium by diet, 2) light exercise, 3) sunbathing, 4) drug administration, and the like have been performed. Calcium salts such as calcium carbonate and calcium phosphate, and natural calcium such as bovine bone meal, eggshell and fish bone powder are used for supplementing calcium by meal. For light exercise, light running and walking are considered to be good, but if the body is weak, light exercise becomes troublesome, and even if a bedridden elderly person, it is almost impossible to exercise. Third sunbathing, but in terms of supplementation of activated vitamin D ₃ it is good, it is not enough this. In the last drug administration, 1
It is known that α-hydroxyvitamin D ₃ , calcitonin preparations and the like are effective for treating and improving osteoporosis. It should be noted that osteoarticular diseases such as rheumatism and periodontal diseases eventually cause bone resorption, and it is considered that these can be improved by suppressing bone resorption.

In order to obtain substances effective for the prevention and treatment of such various bone diseases and osteoarticular diseases and periodontal diseases, the present inventors have proposed an osteoblast growth factor in whey protein and an inhibitor of bone resorption. We have continued to search for factors and fractions with bone strengthening effects. That is, an attempt was made to fractionate milk, particularly whey protein, and fractionate a fraction having an action of inhibiting bone resorption of osteoclasts. As a result, it was found that a protein and peptide mixture obtained by treating a water-soluble fraction in whey protein with a reverse osmosis membrane, an electrodialysis membrane or the like and desalting it has a bone-enhancing effect (Japanese Patent Laid-Open No. -183371). Then, the aqueous solution of this protein and peptide mixture was subjected to ethanol treatment, heat treatment, salt treatment, ultrafiltration membrane treatment, and the like, and it was found that the fraction obtained had a bone strengthening effect (Japanese Patent Application Laid-Open No. H5-176715). JP-A-5-320066). In addition, the present inventors have found that a basic protein present in a trace amount in milk has a collagen synthesis promoting effect in osteoblasts and a bone resorption preventing effect (JP-A-7-20).
No. 7509).

[0004] On the other hand, cystatin, as a cysteine protease inhibitor, is a substance that inhibits the proteolytic activity of a cysteine protease having an SH group at its active center, and is found in animal tissues, cells, blood and urine. In addition, as a useful effect of cystatin, a virus growth inhibitory effect has been confirmed (Biochem. Biophys.
s. Commun., vol. 127, p. 1072, 1985). In recent years, with aging, osteoporosis caused by bone resorption of osteoclasts has been rapidly increasing. At present, calcitonin preparations are known as medicines for suppressing bone resorption of osteoclasts. However, this calcitonin preparation is a hormonal preparation used as a pharmaceutical, and a safe substance that can be used as a food material has not been studied at present. In addition, a large amount of cystatin that can be used as a food material has not been obtained from animal tissues, cells, blood, and urine.

[0005]

DISCLOSURE OF THE INVENTION The present inventors have tried to separate and purify an active substance having a bone resorption inhibiting action for a basic protein having a bone resorption inhibiting action, and identified the separated and purified substance. The material was identified as milk-based cystatin. It has also been found that this milk-based cystatin has an effect of specifically inhibiting bone resorption of osteoclasts, as compared with other species and types of cystatin derived from other than milk (Japanese Patent Application No. 11-110). No. 89946). Under such circumstances, there is a need for a method for producing a large amount of milk-based cystatin that can be used as a food material, at a low cost. Therefore, an object of the present invention is to provide a method for producing a milk-basic cystatin-rich fraction from a milk-derived basic protein. In addition, it has been known that the protease-degraded product of the milk-basic cystatin-rich fraction has a bone resorption inhibiting effect. Providing is also an issue. Furthermore, in the present invention, the prevention and improvement of various bone diseases such as osteoporosis, osteoarticular diseases such as rheumatism, and periodontal disease, in which a milk-basic cystatin-rich fraction or a milk-basic cystatin-rich fraction decomposition product is blended. It is an object to provide food and drink, medicine, or feed for food.

[0006]

According to the present invention, a basic protein composition derived from milk is brought into contact with an anion exchange resin to recover a fraction that does not adsorb to the resin, and then the fraction is brought into contact with a cation exchange resin. The fraction adsorbed on the resin is eluted with an eluate and collected to produce a milk-basic cystatin-containing fraction. In order to obtain a milk-derived basic protein composition as a raw material of the present invention, milk can be used as it is, or reduced milk, skim milk, whey, or the like of such milk can be used. Most of the milk-derived basic protein composition, which is present only in trace amounts in milk, is lactoferrin and lactoperoxidase.After contacting milk with a cation exchange resin, the fraction adsorbed on the resin is salted. By eluting with an eluate having a concentration of 0.1 to 1.0 M,
A large amount of a milk-derived basic protein composition can be obtained. As an eluate for eluting the fraction adsorbed on the cation exchange resin, a solution containing 0.1 to 1.0 M of a salt such as sodium chloride may be used. As the anion exchange resin used in the present invention, commercially available Q Sepharose (manufactured by Pharmacia) and the like are preferable, and as the cation exchange resin, commercially available S Sepharose (manufactured by Pharmacia) and the like are preferable. The present invention also includes producing a milk-basic cystatin-rich fraction hydrolyzate obtained by decomposing a milk-basic cystatin-rich fraction produced by the above method with a protease. Furthermore, in the present invention, a milk-basic cystatin-rich fraction and / or a milk-basic cystatin-rich fraction hydrolyzate produced by the above method are blended, and calcium and / or vitamins are further added as necessary. Blended,
Foods, beverages, medicines or feeds for prevention and improvement of various bone diseases such as osteoporosis, bone joint diseases such as rheumatism, and periodontal disease are also included.

[0007]

According to the method of the present invention, a milk-basic cystatin-rich fraction can be produced in a large amount at low cost from a milk-derived basic protein composition. As milk, raw milk such as cow milk, milk powder, skim milk powder, reduced milk and the like are used. In addition, the milk-basic cystatin-rich fraction hydrolyzate obtained by the method of the present invention is the above-mentioned milk-basic cystatin trypsin, chymotrypsin, pepsin, papain, kallikrein, cathepsin, thermolysin,
It is a peptide mixture that has been limitedly degraded with a protease such as V8 protease. The obtained milk-basic cystatin-rich fraction and the milk-basic cystatin-rich fraction degraded product are preferably concentrated or dried as appropriate. Furthermore, in the present invention, such a milk-basic cystatin-rich fraction and / or a milk-basic cystatin-rich fraction degraded product is blended into foods, drinks, medicines, feeds, or the like to obtain various bone diseases such as osteoporosis and the like. It can be used for prevention and improvement of osteoarticular diseases such as rheumatism and periodontal diseases.
That is, it may be incorporated into foods and drinks such as milk, milk drinks, juices, jellies, biscuits, breads, noodles, sausages and feeds, and may be used as medicines such as tablets and powders, toothpastes, and mouthwashes.

[0008] Various bone diseases such as osteoporosis, osteoarticular diseases such as rheumatism, and periodontal diseases containing a milk-basic cystatin-rich fraction produced by the method of the present invention or a milk-basic cystatin-rich fraction decomposed product. It is preferable to blend calcium having good absorbability into foods, drinks, medicines or feeds for prevention and improvement. Examples of such absorptive calcium include calcium salts such as calcium chloride, calcium carbonate and calcium lactate, natural calcium such as bovine bone meal, eggshell and fish bone meal, and milk-derived calcium compositions. it can. It is also preferable to add vitamins effective for bone, such as vitamin D and vitamin K. The effects of such calcium and vitamins are synergistic because the mechanism of action is different from that of milk-based cystatin and its degradation products. In addition, milk-based cystatin and its decomposed product have high heat stability and extremely excellent properties as a food material. In the present invention, in the case of adults, milk-basic cystatin and its decomposed products are used per day.
50mg ~ 500mg may be taken in several divided doses. As described above, by ingesting milk-based cystatin and its degradation product, various bone diseases such as osteoporosis, bone joint diseases such as rheumatism, and periodontal disease can be prevented and improved.
Next, the present invention will be described with reference to examples.

[0009]

Example 1 A column packed with 3 kg of a sulfonated chitopearl (Fujibo Co., Ltd.) of a cation exchange resin was sufficiently washed with deionized water. After passing 300 l of unsterilized skim milk through the column at a flow rate of 100 ml / min, the column is washed thoroughly with deionized water and then with 0.02 M bicarbonate buffer (pH 6.7) containing 0.87 M sodium chloride. The milk-derived basic protein adsorbed on the resin was eluted and collected. The eluate was desalted and concentrated by reverse osmosis membrane treatment, and then lyophilized to obtain a powdered milk-derived basic protein composition. This operation was performed five times to obtain 1.65 kg of a milk-derived basic protein composition.
A solution prepared by dissolving 1 kg of the milk-derived basic protein composition in a sodium carbonate buffer (pH 9.0) to a concentration of 1% by weight is passed through a column packed with Q Sepharose as an anion exchange resin (20 l). Per hour), and 100 l of sodium carbonate buffer (pH 9.0) was passed through to obtain 200 l of a fraction not adsorbed to the resin. Next, the obtained fraction was passed through a column packed with S-Sepharose as a cation exchange resin (20
l / h), sodium carbonate buffer (pH 9.0) 201 and
After the column was washed by passing a 0.3 M aqueous sodium chloride solution 151 in sequence, 10 l of a 0.5 M aqueous sodium chloride solution was passed to obtain 10 l of an eluted fraction. The eluted fraction was concentrated by ultrafiltration (UF) (10 kDa cut in molecular weight) until the concentration became 10 times, and further desalted by diafiltration (DF) to obtain 1 kg of a concentrated desalted fraction. . The concentrated desalted fraction was lyophilized to obtain 74 g of a milk-basic cystatin-rich fraction. The milk-basic cystatin-rich fraction contained 1.6% by weight of milk-basic cystatin.

[0010]

Example 2 1 g of the milk-basic cystatin-rich fraction obtained in Example 1 was suspended in 100 ml of water, and to this suspension was added pancreatin to a final concentration of 1% by weight.
The enzyme reaction was performed at 37 ° C. for 5 hours. After heating at 90 ° C. for 5 minutes to inactivate the enzyme, the solution was freeze-dried to obtain 0.95 g of a milk-basic cystatin-rich fraction hydrolyzate.

[0011]

[Test Example 1] The long bones of 10-20 day old ICR mice were excised, soft tissues were removed, and the long bones were mechanically sliced in an α-MEM solution containing 5% fetal bovine serum. The whole bone marrow cells including osteoclasts were obtained by cutting. About these cells, about 2 million cells were spotted on an ivory piece with an α-MEM solution containing 5% fetal calf serum. After several hours, 5% after adding the test sample
An α-MEM solution containing fetal calf serum was added and cultured for 3 days, and the bone resorption activity of osteoclasts was examined. The bone resorption activity was evaluated by removing the cells on the ivory piece after culture, staining with hematoxylin, analyzing the image with PIASLA-555,
it). The milk-basic cystatin-rich fraction (test sample A) obtained in Example 1 and the milk-basic cystatin-rich fraction degraded product (test sample B) obtained in Example 2 were each 500 mg / mg.
A test sample solution having a concentration of ml was prepared, and the bone resorption activity was examined. That is, the number of bone resorption pits (pits) when the cells were cultured with the test sample added was counted, and the number of bone resorption pits (pits) when the cells were cultured without the addition of the test sample was 100%. The absorption activity is shown in Table 1.

[0012]

[Table 1]

From Table 1, it was found that the milk-basic cystatin-rich fraction and the hydrolyzate of the milk-basic cystatin-rich fraction had an effect of inhibiting bone resorption activity.

[0014]

Test Example 2 An animal experiment was performed on the test sample A obtained in Example 1 and the test sample B obtained in Example 2 using an osteoporosis model rat. Table 2 shows the basic composition of the feed administered to the rats. The calcium: phosphorus ratio was set to 1: 1 so that the amount of calcium and phosphorus in the feed was 300 mg per 100 g of feed in all groups.

[0015]

[Table 2] 蔗 Sucrose 50.0 (% by weight) Casein 20.0 Corn starch 15.0 Cellulose 5.0 Corn oil 5.0 Vitamin mixture (including choline) 1.0 Mineral mixing 4.0 ¹⁾ ──────────────────────── 1) Calcium carbonate was used as the calcium source.

[0016] Test samples A or B were added to a feed having the basic composition shown in Table 2 to prepare the following test meals. Test meal 1: feed with basic composition shown in Table 2 + test sample A
(0.05mg / 100g) Test meal 2: Feed with basic composition shown in Table 2 + Test sample B
(0.05 mg / 100 g) Test meal 3: Feed having the basic composition shown in Table 2 (however, calcium derived from milk (JP-A-4-306622) was used instead of calcium carbonate as a calcium source) + test sample A (0.05mg / 100g)

The animals used were SD female rats of 40 weeks of age. The osteoporosis model rat was prepared by pre-breeding for one week, performing an ovariectomy operation, and breeding on a low calcium diet for two months. At that time, a sham rat without ovaries was prepared by performing sham operation. Each test group was divided into seven groups, and each test meal was administered for one month. The sham group and the control group were administered a feed having the basic composition shown in Table 2. After the administration of the test meal, the femurs of the rats in each test group were excised, the amount of bone mineral was measured with a bone mineral amount measuring device, and the bone strength was measured with a fracture characteristic measuring device. The test results are shown in Tables 3 and 4.

[0018]

[Table 3] ──────────────────────── Test group Bone mineral content (mg, ± SD) ─────────── ───────────── Sham group 138.2 ± 3.6 Control group 89.1 ± 3.2 Test food 1 administration group 101.2 ± 3.9 Test food 2 administration group 113.3 ± 3.1 Test diet 3 administration group 110.1 ± 3.5 ─── ─────────────────────

As shown in Table 3, the amount of bone mineral in the femur was
The test food administration group showed a statistically significantly higher value than the control group. From this, it was found that the test samples A and B have a bone resorption preventing action. It was also found that the addition of milk-derived calcium with good absorbability further enhanced the action.

[0020]

[Table 4] ────────────────────── Test group Bone breaking force (10 ⁶ dyn) ────────────── ─────── Sham group 13.4 ± 2.3 Control group 6.5 ± 2.1 Test meal 1 group 8.6 ± 2.2 Test meal 2 group 11.1 ± 2.0 Test meal 3 group 10.5 ± 2.4 ───────── ─────────────

As shown in Table 4, the bone breaking strength was statistically significantly higher in the test food administration group than in the control group.
From this, it was found that Test Samples A and B had a bone strengthening effect. It was also found that the addition of milk-derived calcium with good absorbability further enhanced the action.

[0022]

Test Example 3 A test sample A (0.05 mg / 100 g) and vitamin D (200 IU) were added to an aqueous solution having the basic composition shown in Table 5, mixed, filled in a container, and then sterilized by heating and the beverage ( Test article). In addition, the drink (control product) which added albumin (0.05 mg / 100g) instead of the test sample A was manufactured similarly.

[0023]

[Table 5] 結晶 Crystal glucose 15.0 (% by weight) Calcium 0.5 Water 74.5 ───────────── ─────────

Patients with osteoarthritis (shrinkage of joint dehiscence)
Twenty people were divided into two groups of 10 people, and each of the above drinks was drunk for one month, and the amount of urinary deoxypyridinoline, which is a bone metabolic marker of bone resorption before and after drinking, was measured. Also,
Subjective symptoms were confirmed by interview. The test results are shown in Tables 6 and 7.

[0025]

[Table 6] 減少 Decrease in deoxypyridinoline (mg, ± SD) ──── ────────────────────────── Control product administration group 0.25 ± 0.2 Test product administration group 0.56 ± 0.2 ────────── ────────────────────

As shown in Table 6, although the amount of deoxypyridinoline was reduced in the control group to which calcium and vitamin were added, the amount of deoxypyridinoline was further reduced in the test group. From this, it was found that the test sample A extremely suppressed the bone resorption due to the destruction of the bone.

[0027]

[Table 7] 数 Number of patients with joint pain before and after administration (persons)投 与 Control product administration group Test product administration group Before / after / after / after ─────────────────関節 Joint pain due to physical compression 10 10 10 6 Joint pain during operation 6 5 5 2 Joint pain at bedtime 5 6 5 1 Joint pain during fatigue 9 8 8 4 Weakness 6 5 7 5 Joint pain throughout laceration 9 8 9 6 ────────────────────────────────

As shown in Table 7, it can be seen that the pain was reduced in each item also for the joint pain.

[0029]

[Test Example 4] A 6-week-old golden hamster was preliminarily reared for 1 week, and then sterilized surgical suture silk thread No. 4 was wound around the cervical region of M1 five times under ether anesthesia. : Keyes, PH and Jordan: Archs. Oral.Biol, vo
l.9, pp.377-400, 1964) caused periodontal disease. A group consisting of 18 hamsters was used as a test group.
Each time, a treatment for continuously immersing the oral cavity for about 10 minutes was performed. In addition,
The control group was treated with distilled water. Four weeks after the start of the treatment, a fixed perfusion was performed for about 20 minutes using a 2.5% glutaraldehyde solution (pH 7.4), and both sides of the mandible were removed. To evaluate the amount of alveolar bone loss, fixation with a 2.5% glutaraldehyde solution was performed and a soft X-ray photograph was taken using an image analyzer (PI
AS LA-555) was analyzed, and the area between the enamel cement boundary near M1 and the alveolar bone crest was measured to evaluate the alveolar bone loss.
Table 8 shows the test results.

[0030]

[Table 8] ───────────────────────────────── Treatment Control group Test group A Test group B (Use sample A) (Use test sample B) ───────────────────────────────── Reduced area (mm ² ) 4 days 0.32 0.25 ^* 0.21 ^* 9 days 0.89 0.65 ^* 0.53 ^* ───────────────────────────────── ^* control Significantly different from group (P <0.05)

According to Table 8, the amount of alveolar bone loss was significantly lower in the test group than in the control group, and this effect was concentration-dependent. This indicates that the milk-basic cystatin-rich fraction and its decomposed product suppress the reduction of alveolar bone and are effective in preventing periodontal disease.

[0032]

Example 3 Prevention and improvement of various bone diseases such as osteoporosis and various osteoarticular diseases such as rheumatism and various bone diseases such as osteoporosis and rheumatism by mixing each component with the composition shown in Table 9 to prepare a dough, molding and doughing. Biscuits were manufactured.

[0033]

[Table 9] ────────────────────── Flour 50.0 (% by weight) Sugar 20.0 Salt 0.5 Margarine 12.5 Egg 12.1 Water 4.0 Sodium bicarbonate 0.1 Ammonium bicarbonate 0.2 Calcium carbonate 0.5 Test sample A 0.1 ──────────────────────

[0034]

Example 4 Jelly for preventing and improving various bone diseases such as osteoporosis and various osteoarticular diseases such as rheumatism and various bone diseases such as osteoporosis and rheumatism after mixing each component with the composition shown in Table 10 and filling in a container. Was manufactured.

[0035]

[Table 10] ──────────────────── Fructose 20.00 (% by weight) Granulated sugar 15.00 starch syrup 5.00 Agar 1.00 Test sample B 0.01 Flavor 0.10 Calcium carbonate 0.10 Water 58.79 ── ──────────────────

[0036]

Example 5 The components shown in Table 11 were mixed to prepare a cheese at an emulsification temperature of 85 ° C. for preventing and improving various bone diseases such as osteoporosis and various osteoarticular diseases such as rheumatism.

[0037]

表 Gouda cheese 43.0 (% by weight) Cheddar cheese 43.5 Sodium citrate 2.0 Test sample B 0.1 Calcium derived from milk 1.0 Water 10.4 ─────────────────────

[0038]

Example 6 After 12% skim milk was heat-sterilized at 90 ° C. for 20 minutes, the skim milk was inoculated with Lactobacillus acidophilus or L. acidophilus, and S. thermophilus was inoculated. Two types of starter culture were prepared. Then, using a yogurt mix containing milk as a main component, each component is mixed with the composition shown in Table 12, fermented and cooled in the usual manner, and various bone diseases such as osteoporosis and various bone joint diseases such as rheumatism. Yogurt for the prevention and improvement of corn.

[0039]

１２ Yogurt mix 96.99 (% by weight) L. acidophilus 1.50 S. thermophilus 1.50 Test sample A 0.01 ──── ───────────────────

[0040]

Example 7 The components shown in Table 13 were mixed together and pressed to produce tablets for preventing and improving various bone diseases such as osteoporosis and various osteoarticular diseases such as rheumatism.

[0041]

[Table 13] 水 Hydrous crystalline glucose 93.50 (% by weight) Test sample A 0.05 Calcium 5.00 Vitamin D (200 IU) Sugar Ester 1.00 Fragrance 0.45 ──────────────────────────

[0042]

Example 8 The ingredients shown in Table 14 were mixed to prepare a dog breeding feed (dog food) for preventing and improving various bone diseases such as osteoporosis and various osteoarticular diseases such as rheumatism.

[0043]

[Table 14] ──────────────────── Soybean meal 12.00 (% by weight) Skim milk powder 14.00 Soybean oil 4.00 Corn oil 2.00 Palm oil 27.99 Corn starch 15.00 Flour 9.00 Bran 2.00 Vitamin mixture 9.00 Mineral mixture 2.00 Cellulose 3.00 Test sample A 0.01 ────────────────────

[0044]

Example 9 Each component was mixed according to the composition shown in Table 15 to prepare a cream. The cream was filled in a container to produce a dentifrice for preventing and improving periodontal disease.

[0045]

Glycerin 70.49 (% by weight) Silicon dioxide 20.00 Xanthan gum 1.00 Mint flavor 1.00 Titanium dioxide 0.70 Sodium fluoride 0.30 Distillation Water 6.50 Test sample B 0.01 ────────────────────────

[0046]

According to the present invention, a milk-derived basic protein composition is brought into contact with an anion exchange resin to collect a fraction that does not adsorb to the resin, and then the fraction is contacted with a cation exchange resin to adsorb to the resin. By eluting the fraction thus obtained with an eluate, a high-concentration milk-basic cystatin-rich fraction can be produced. Further, by decomposing this fraction containing milk-basic cystatin high with a protease, it is possible to produce a decomposition product of the fraction containing milk-basic cystatin high. The obtained milk-basic cystatin-rich fraction and the cystatin-rich fraction degraded product are blended with foods, drinks, medicines, feeds, etc., to obtain various bone diseases such as osteoporosis, osteoarthritis such as rheumatism, and periodontal disease. It is useful for prevention and amelioration of disease.

Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat II (reference) A23L 1/30 A23L 1/30 A 4C087 1/305 1/305 4C206 2/52 2/38 P 2/66 A61K 31 / 122 2/38 31/59 A61K 31/122 33/06 31/59 35/20 33/06 45/00 35/20 A61P 1/02 38/55 19/02 45/00 19/10 A61P 1/02 43 / 00 111 19/02 A23L 2/00 F 19/10 J 43/00 111 A61K 37/64 (72) Inventor Hiroshi Kawakami 204-5 Fujima, Kawagoe-shi, Saitama F-term (reference) 2B150 AB03 AB10 AE26 BB04 CC11 DA47 DD01 DE01 DF10 DH04 4B017 LC03 LK01 LK15 LK16 LK18 LK23 LL09 LP01 LP06 LP08 4B018 LB01 LB07 MD04 MD20 MD23 MD71 ME05 ME09 MF01 MF12 4C084 AA02 AA03 AA17 CA38 DC02 NA14 ZA671 ZA961AAMA MA ZA961 MA04 4C087 BB39 NA14 ZA67 ZA96 ZA97 ZC20 4C206 AA01 CB28 KA04 MA01 MA03 MA04 MA72 NA14 ZA67 ZA96 ZA97 ZC20

Claims (4)

[Claims] 1. A milk-derived basic protein composition is contacted with an anion exchange resin to collect a fraction that does not adsorb to the resin, and then the fraction is contacted with a cation exchange resin to separate the fraction adsorbed to the resin. A method for producing a milk-basic cystatin-rich fraction, characterized in that the fraction is eluted and recovered with an eluate. 2. A method for producing a milk-basic cystatin-rich fraction degraded product, comprising decomposing a milk-basic cystatin-rich fraction produced by the production method according to claim 1 with a protease. 3. Various types of osteoporosis and the like, comprising a milk-basic cystatin-rich fraction and / or a milk-basic cystatin-rich fraction degraded product produced by the production method according to claim 1 or 2. Food and drink, medicine or feed for prevention and improvement of bone diseases and osteoarticular diseases such as rheumatism and periodontal diseases. 4. The food or drink, medicine or feed according to claim 3, further comprising calcium and / or vitamin.