Bone resorption suppressing agent

Drug – bio-affecting and body treating compositions – Preparations characterized by special physical form – Food or edible as carrier for pharmaceutical

Reexamination Certificate

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C424S400000, C424S440000, C424S451000, C424S464000

Reexamination Certificate

active

06607743

ABSTRACT:

BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to bone resorption suppressing agents comprising a milk-derived basic cystatin and/or milk-derived basic cystatin decomposition product as an effective component, and in particular to bone resorption suppressing agents to be used for the prevention and treatment of bone joint diseases or periodontal diseases.
The present invention also relates to drinks, food products and feed in which a milk-derived basic cystatin and/or milk-derived basic cystatin decomposition product are admixed to provide an activity to prevent and treat bone joint diseases or periodontal diseases. In particular, the present invention relates to drinks, food products and feed to which a highly absorbable calcium composition and vitamin D and/or vitamin K, in addition to a milk-derived basic cystatin and/or milk-derived basic cystatin decomposition product, are admixed to provide an activity to prevent and treat bone joint diseases or periodontal diseases.
2. Description of the Related Art
In recent years, the number of people suffering from bone joint diseases such as osteoporosis and rheumatism has been on the rise because of the aging population. Bone formation and bone resorption continuously take place in bone tissue and are well balanced in early life, but the balance is lost with an increase in bone resorption with aging for various reasons. If this unbalance continues for a long period of time, the bone tissue becomes fragile, which results in bone diseases such as osteoporosis, bone fractures and lumbago. It is believed that if this uncoupling can be prevented, osteoporosis, bone fractures, lumbago or the like can be prevented.
Conventional methods to prevent and treat bone diseases by preventing the uncoupling include (1) dietary supplementation of calcium, (2) moderate exercise, (3) sunbathing, and (4) therapy by medicines.
For dietary supplementation, calcium salts such as calcium carbonate and calcium phosphate, natural calcium supplements such as bovine bone powders, egg shells and fish bone powders are used. To date, such food products and food materials have been used exclusively for calcium supplementation.
For moderate exercise, moderate running or walking are highly recommended. However, even moderate exercise is difficult for people who are physically weak and, not to mention, almost impossible for elderly people who are confined to bed.
Sunbathing is good for the supplementation of activated vitamin D
3
, but not sufficient by itself.
As for therapeutic medicines, 1&agr;-hydroxy vitamin D
3
, calcitonin preparations or the like are known to be effective to cure and treat osteoporosis. Calcitonin preparations are medicinal hormone preparations. Safe substances for calcitonin obtainable from food materials have not been presently studied. Calcitonin is difficult to prepare in bulk and to provide as a safe food material since animal tissues, cells, blood, or urine have to be used as raw materials.
It is believed that rheumatism, a joint disease, can be prevented and treated by suppressing bone resorption since it is associated with the bone resorption.
In recent years, periodontal diseases have also become a serious social problem. Unlike dental caries, periodontal diseases weaken the roots of teeth, which makes even healthy teeth useless. Today, many people develop symptoms of periodontal diseases. Periodontal diseases can be more serious than dental caries.
Today, means to prevent periodontal diseases are to prevent the growth of causative microorganisms, for example, the removal of dental plaque or gargling using mouthwashes containing antibacterial agents or the like. However, these means seem to be less effective for highly developed symptoms. Namely, in the late stage of periodontal diseases, the alveolar bone mass decreases, and once alveolar bones are lost, unregenerable symptoms occur. Teeth loss due to periodontal diseases then makes eating difficult and painful, which is disturbing in everyday life. Thus, there is a need for effective means for the prevention and treatment of periodontal diseases.
However, so far, there is no agent available for the prevention and treatment of periodontal diseases which effectively suppresses a decrease in alveolar bone mass.
Thus, like osteoporosis, periodontal diseases have become a serious social problem. Therefore, an effective treatment for periodontal diseases is will greatly contribute to people's health, and accordingly is necessary,.
The present inventors intensively searched for a milk whey fraction having osteoblast growth stimulating activity, bone resorption suppressing activity and bone strengthening activity, in order to obtain a material which can be used for the prevention and treatment of bone junction diseases and periodontal diseases. Namely, the present inventors fractionated proteins in milk, in particular milk whey, in an attempt to obtain a fraction having a bone resorption suppressing activity, and found a bone strengthening activity in a protein-peptide mixture which was obtained by treating a water soluble fraction of whey proteins with a reverse osmotic membrane or electrodialysis to remove whey-derived salts (Japanese Patent Laid-open No. 4-183371). Furthermore, the present inventors found that a fraction obtained by treating an aqueous solution of this protein-peptide mixture with ethanol, heat, salts or an ultrafiltration membrane has a bone strengthening activity (Japanese Patent Laid-open No. 5-176715, Japanese Patent Laid-open No. 5-320066). The present inventors also found that basic proteins present in milk in trace amounts have an osteoblast collagen synthesis stimulating activity and bone resorption suppressing activity (Japanese Patent Laid-open No. 8-151331).
Cystatin is a cysteine protease inhibitor which inhibits proteolytic activity of cysteine proteases having an SH group in the active center and is found in animal tissues, cells and urine. A virus growth inhibiting activity was recognized as a useful activity of cystatin (Biochem. Biophys. Res. Commun., Vol. 127, p. 1072, 1985).
Japanese Patent Laid-open No. 2-223529 describes the use of cystatin as anti-allergic agents and as therapeutic agents for bone diseases in the form of an injectable preparation, suppository, nasal powder or the like. More specifically, it described a test result wherein the blood calcium level was reduced in rats by intravenously injecting rats with rat-derived cystatin prepared by genetic engineering. However, one cannot readily conclude that cystatin has an activity to prevent and treat bone joint diseases such as osteoporosis and rheumatism, solely from this result.
Furthermore, until now, a bone resorption suppressing agent comprising a milk-derived basic cystatin and/or milk-derived basic cystatin decomposition product which is effective by administration orally but not necessarily intravenously, as an effective component, are not known. Also, a drink, food product or feed which contains such components in higher concentrations, along with calcium and vitamins, and can be administered orally is not known.
Although the present inventors previously applied for a patent (Japanese Patent Laid-open No. 7-126294) on a cysteine protease inhibitor other than the milk-derived basic cystatin, the bone resorption suppressing activity of the milk-derived basic cystatin of the present invention is higher than that of said inhibitor.
SUMMARY OF THE INVENTION
The present inventors tried to isolate and purify an active substance from of basic protein fraction having a bone resorption suppressing activity, identified the resulting substance, and found this substance is a milk-derived basic cystatin. Furthermore, the present inventors found that the milk-derived basic cystatin has a particularly high bone resorption suppressing activity as compared with cystatins from other origins. Further, the present inventors found that a decomposition product of the milk-derived basic cystatin also has a high bone resorption suppressing activity, and thus compl

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