JP2005035963A - Bone, rheumatism and periodontal disease-improving agent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a therapeutic agent excellent in treatment or improvement of metabolic bone diseases such as osteoporosis caused by decrease of bone salt, having bone-improving action, rheumatism-improving action and periodontal disease-improving agent. <P>SOLUTION: The present inventor discovered that limitin has bone-improving action, rheumatism-improving action and periodontal disease-improving action. The present invention provides a bone amount-improving and bone-strengthening agent, a rheumatism-improving agent and a periodontal disease-improving agent each containing limitin as an active ingredient. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

Detailed Description of the Invention

  The present invention relates to a novel bone mass improving and bone strengthening and rheumatic agent. More specifically, the present invention relates to a bone mass improving and bone strengthening agent, rheumatism improving agent and periodontal disease containing limitine as an active ingredient.

  In recent years, bone and joint diseases such as osteoporosis and rheumatism are increasing with aging. In bone tissue, bone formation and bone resorption are constantly carried out. When young, bone formation and bone resorption are balanced, but the balance tends to bone resorption due to various causes with aging. If this state continues for a long period of time, the bone tissue becomes brittle and causes bone diseases such as osteoporosis, fractures, and back pain. If this uncoupling can be prevented, it is believed that osteoporosis, fractures, back pain and the like can be prevented.

  Conventionally, (1) dietary calcium supplementation, (2) light exercise, (3) sunbathing, (4) drug treatment, and the like have been performed as methods for preventing uncoupling and preventing / treating bone diseases.

  For calcium supplementation with meals, calcium salts such as calcium carbonate and calcium phosphate and natural calcium preparations such as cow bone meal, eggshell and fish bone meal are used. All previous uses for food and food ingredients are calcium supplements.

  For light exercise, light running and walking are good, but if the body is weak, light exercise becomes troublesome, and even if you are a bedridden elderly person, you can hardly exercise.

Further, sunbathing but there is a good in terms of supplementation of activated vitamin D _3, it is not enough this.

As drug treatment, it is known that 1α-hydroxyvitamin D ₃ , calcitonin preparation and the like are effective for the treatment and improvement of osteoporosis. The calcitonin preparation is a hormone preparation as a pharmaceutical product. However, there are still few effective drugs that work directly on osteoclasts to stop bone loss.

  On the other hand, since rheumatism, which is a joint disease, is a disease accompanied by bone resorption, it is considered that rheumatism can be prevented and improved by suppressing bone resorption.

  In recent years, periodontal disease has become a major social problem. Periodontal disease, unlike caries, is a disease that weakens the root of the tooth and makes it impossible to use non-toothed teeth. Recently, many people have shown signs of periodontal disease, rather than a caries disease.

  Currently, the prevention of periodontal disease is from the viewpoint of preventing the growth of microorganisms that cause causative bacteria, such as removing plaque and gargles using antibacterial agents (mouthwash). Although prevention has been carried out, these methods are considered to be less effective against symptoms that have progressed excessively. That is, at the end of periodontal disease, the alveolar bone is decreased, and once the alveolar bone is lost, it is difficult to regenerate.

  In addition, when teeth are lost due to periodontal disease, it becomes difficult to eat foods, and because it causes pain, etc., there is a need for effective preventive and therapeutic means. However, at present, there is no preventive and ameliorating agent for periodontal disease having an effect of suppressing the decrease in alveolar bone.

Problems to be solved by the invention

  In this way, bone diseases, rheumatic diseases, and periodontal diseases have become major social problems, and if this also has an effect on improvement, it is considered that there are significant benefits to people's health. There is also a great need for improvements. Finding an active ingredient having a new and effective action for solving such a problem is a problem to be solved. That is, an object of the present invention is to provide a therapeutic agent having an excellent bone improving action, rheumatism improving action and periodontal disease improving action for metabolic bone diseases such as osteoporosis caused by bone mineral loss.

Means for solving the problem

  As a result of the sincere efforts, the present inventors have found that Limitin has a bone improving action, a rheumatism improving action, and a periodontal disease improving action, and has led to the present invention. That is, a bone mass improving and bone strengthening agent containing limitine as an active ingredient, a rheumatism improving agent containing limitine as an active ingredient, and a periodontal disease improving agent containing limitine as an active ingredient.

Limitin is 30% structural homology in mice as an interferon (hereinafter abbreviated as IFN) -like protein, but the substance is different from IFN. It also binds to the IFN receptor and activates IFN modulators. IFN is a substance produced by various factors including viral infection, and is a kind of cytokine having biological activity such as antiviral action, antitumor action, and immunoregulatory action. IFN includes α-type, β-type, and γ-type.
As the limitin used in the present invention, either naturally derived or genetically modified can be used.

  In the bone mass increasing agent of the present invention, limitin may be used alone or in combination with other bone mass increasing agents, bone resorption promoting factor inhibitors, rheumatism improving agents, periodontal disease improving agents, and the like. Also good. As a combination method, for example, a combination form may be used, or a preparation composed of a single agent may be used in combination.

  The bone mass increasing agent of the present invention can be administered orally or parenterally, for example, prepared in the form of injections, tablets, capsules, granules, pills, troches, powders, liquids, etc. It may contain a pharmacologically acceptable medium suitable for the dosage form. In the case of injections, pH buffers, preservatives, stabilizers, solubilizers, sterile isotonic saline, local anesthetics and the like are generally used. Moreover, formulation is performed according to the formulation of a well-known injection or instillation, For example, buffer agents, such as a trisglycine buffer, can be used.

  As the dose of limitin used in the present invention, it is desirable to use an amount corresponding to the symptom, and it is generally possible to use about 10 ng / day to about 10 mg / day. Preferably, 1 μg / day to 100 μg / day is used.

  In general, it is desirable to administer until an effect is observed, and it is preferably 2 to 12 weeks. In addition, any mode of continuous administration or intermittent administration within the administration period is possible, and they can be appropriately combined.

  As a method for administering the bone mass increasing agent of the present invention, a generally known embodiment is possible. In the case of an injection, it is generally administered by intramuscular injection, intravenous injection or subcutaneous injection. It is also possible to administer an IFN in a sustained release formulation. For example, the bone mass increasing agent of the present invention can be applied to the oral cavity, pharynx, etc. by dissolving or disintegrating gradually in the mouth in the form of a film.

  Hereinafter, examples and experimental examples will be given to describe the present invention in more detail, but the present invention is not limited to these examples.

  Example 1 Limitin was obtained according to Oritani K et al (Nat. Med. 6, 659-666).

Test example 1

Bone resorption inhibitory effect of limitine After removing the long bones of 10-20 days old ICR mice and removing soft tissues, the bones were mechanically minced in an α-MEM solution containing 5% fetal bovine serum. And whole bone marrow cells containing osteoclasts were obtained. About 2 × 10 6 cells were spotted on an ivory piece with an α-MEM solution containing 5% fetal calf serum. Several hours later, an α-MEM solution containing 5% fetal bovine serum to which milk-derived basic cystatin or a milk-derived basic cystatin degradation product was added to a concentration of 50 ng / ml was added at 37 ° C. in the presence of 5% CO 2. Cultivate for 5 days and examine osteoclastic bone resorption activity.
After culturing, the cells on the ivory pieces were peeled off and stained with hematoxylin, and image analysis was performed with an image analyzer (PIASLA-555, manufactured by PIAS) to measure the number of bone resorption pits. From the measurement results, the bone resorption activity (%) defined by the following formula was obtained, and the bone resorption inhibitory effect was evaluated.
Bone resorption activity (%)
= (Number of bone resorption pits in the test sample addition group / Number of bone resorption pits in the test sample addition group) × 100
Limitine was used as a test sample. Bone resorption was suppressed in a concentration-dependent manner by adding limitin. The results are shown in FIG.

Test example 2

The bone mass and the bone strengthening effect were examined by the administration test using an osteoporosis model rat of limitin. A feed having an AIN-76 composition was used as a feed for the administration test.
The amount of calcium and phosphorus in the feed was adjusted to 300 mg per 100 g of the co-feed, and the calcium: phosphorus ratio was 1: 1. As experimental animals, SD female rats of 32 weeks of age were used. This rat was preliminarily bred for one week and then subjected to ovariectomy and bred for 2 months on a low calcium diet to create an osteoporosis model rat.
In the administration test, the rats were divided into a control group and a limitin administration group of 6 groups (1 test group: 7 animals), and limitin was administered orally or intravenously for 1 month. As an oral product, one dose of limitin was dissolved in appropriate amounts of 2% atelocollagen aqueous solution, 5% gelatin aqueous solution, and 0.5% glucose, and a freeze-dried film-form oral preparation was obtained.
After administration of the test feed, the femurs of the rats of each test group were removed, the bone mineral content was measured with a bone mineral content measuring device, and the bone strength was measured with a fracture characteristic measuring device. The results are shown in FIGS.

  As shown in FIG. 2, the bone mineral content of the femur was higher in the group in which limitin was orally or intravenously injected than in the control group. Furthermore, as shown in FIG. 3, the femoral fracture force was statistically significantly higher in the limitin group than in the control group. From this, it was found that limitin has a bone strengthening action. From the above results, it was found that limitin has an effect of suppressing bone resorption and an effect of strengthening bone.

Test example 3

Twenty patients with osteoarthritis (joint cleavage contraction) were divided into two groups of 10 people, and the oral film obtained in Test Example 2 was taken for 1 month to improve joints before and after drinking I investigated. The results are shown in Table 4. As shown in FIG. 4, it was found that pain was reduced for each item with respect to joint pain.

Test example 4

  Twenty-four six-week-old golden hamsters were preliminarily raised for one week, and then sterilized surgical suture silk No4 was wrapped around the M1 tooth neck under ether anesthesia five times, and the feed from Keyes et al. (D # 2000: Keyes, P. H. and Jordan: Archs. Oral. Biol. 9: 377-400, 1964), periodontal disease was caused. Next, the golden hamster was divided into 4 groups, and the treatment of continuously immersing the oral cavity for about 10 minutes with a test solution in which 5 μg of each fraction was appropriately diluted was continuously performed twice a day. Seven days after the start of treatment, 6 animals were selected for each group, fixed and perfused with a 2.5% glutaraldehyde solution (pH 7.4) for about 20 minutes, then both sides of the mandible were removed, and the alveoli were excised in the following manner. Bone loss was assessed. That is, both sides of the extracted mandible were fixed with a 2.5% glutaraldehyde solution, then soft X-rays were taken, and the photograph was analyzed using an image analyzer (PIASLA-555), and the enamel cement boundary near M1 And the amount of alveolar bone loss was evaluated by measuring the area between the alveolar bone crests.

  The results are shown in FIG. The golden hamster in the limitin group had significantly lower alveolar bone loss than the control group. From this, it was found that limitin is effective in preventing and improving periodontal disease by suppressing alveolar bone loss.

The invention's effect

The bone and rheumatic agent of the present invention acts to suppress bone resorption and increase bone mass, and is effective in improving rheumatism. Therefore, the bone and rheumatic agent of the present invention is effective in the treatment of metabolic bone diseases such as osteoporosis caused by bone mineral loss. It is useful for treatment and prevention, and for improvement of rheumatic diseases. Furthermore, it is effective in preventing periodontal disease.

“It is a diagram showing the relationship between limitin and bone resorption activity.” “It is a diagram showing the relationship between limitine and bone mineral content.” “It is a diagram showing the relationship between limitin and bone fracture energy.” “It is a figure showing the relationship between the number of joint pain patients (persons) before and after administration.” “It is the figure which showed the relationship between limitin and the amount of decrease in alveolar bone immunity.”

Claims (3)

Bone mass improvement and bone strengthening agent containing limitine as an active ingredient Rheumatoid improving agent containing limitine as an active ingredient Periodontal disease improving agent containing limitine as an active ingredient

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