KR102140493B1 - Composition for preventing or treating of bone diseases comprising denatonium compound - Google Patents

Abstract

The present invention relates to a composition for preventing or treating bone diseases including denagonium as an active ingredient, and particularly, to a pharmaceutical composition or health-aid food for preventing or treating bone diseases including denatonium compound or a salt thereof as an active ingredient. According to the present invention, denatonium or a salt thereof has an excellent activity of inhibiting excessive formation or differentiation of osteoclast, and can effectively improve (i) loss of spongy bone; (ii) a decrease in bone mineral density (BMD), volume of spongy bone and cortical bone (BV), volume ratio of spongy bone and cortical bone (BV/TV), spongy bone thickness (Tb, Th), spongy bone number (Tb, N), spongy bone volume (Tb, V), cortical bone volume (Ct, V) and in cortical bone mineral density (Ct, BMD); and (iii) a condition of inhibiting formation of a vertebra, generated upon the occurrence of bone diseases. Therefore, the composition can be useful for a medicine and health-aid food for preventing, alleviating and treating bone diseases.

Description

Composition for preventing or treating bone diseases comprising denatonium compound

The present invention relates to a composition for the prevention or treatment of bone diseases comprising a denatonium compound or a salt thereof as an active ingredient.

The bone is a calcified connective tissue that is associated with muscles, and the surface is a thick, hard calcified tissue that acts as a support for body balance and protects organs. The inside of the bone is the bone marrow tissue, which is the center of calcium metabolism. Bone is composed of organic substances such as collagen, osteocalcin and osteonectin, and inorganic substances such as calcium, phosphorus and fluorine. Bone formation and bone resorption are always occurring in the living body. Bone formation is promoted by a small amount of parathyroid hormone, androgen, estrogen, fluorine, phosphorus, etc., and bone resorption is promoted by physical decompression, weightlessness, a large amount of parathyroid hormone, and corticosteroids. Therefore, the balance between them is important for bone metabolism. Therefore, bone homeostasis is continuously controlled by bone resoprtion by osteoclasts and bone remodeling by the equivalent action of bone formation by osteoblasts. do.

It is known that two types of cells are involved in bone remodeling, that is, the remodeling process, one of which is an osteoblast that produces bone, and the other is an osteoclast that destroys bone. . However, the imbalance of these two cells, especially the excessive decomposition of bone by excessive formation or activation of osteoclasts, is a major cause of various bone diseases including osteoporosis.

In particular, osteoclastogenesis (osteoclastogenesis) is caused by stimulation of cytokines such as machrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor-κB ligand (RANKL), and osteoclast progenitor cells fuse to each other to multi-nucleate. This is the process by which osteoclasts are formed. The mature osteoclasts cause the cytoskeleton process, and the osteoclast acts as a large ring of osteoclasts to separate the extracellular space from the bone-absorbing space while the osteoclast attaches to the bone. It refers to the process of organizing with (ring) to absorb bone.

Therefore, inhibiting the osteoclast differentiation, maturation, or activity by inhibiting the bone-absorption action may be an effective method of developing various bone disease therapeutic agents.

To date, several substances have been developed to treat bone diseases. Among them, estrogen, which is most frequently used as a therapeutic agent for osteoporosis, has yet to be verified for its actual efficacy and has a disadvantage that it must be continuously taken throughout life, and has a side effect of increasing breast cancer or uterine cancer when administered for a long time. Alendronate, a bisphosphonate-based drug, has a problem in that its efficacy is not clear, absorption in the digestive tract is slow, and inflammation occurs in the gastrointestinal tract and esophageal mucosa. Calcium preparations are known to have excellent effects with few side effects, but they are more prophylactic than therapeutic. In addition, vitamin D preparations such as calcitonin are known, but research on efficacy and side effects has not been sufficiently conducted.

Therefore, there is a need to develop a new therapeutic agent for bone disease with few side effects and excellent effect.

Republic of Korea Registered Patent No. 10-1305555

Accordingly, the present inventors, while studying to develop a new therapeutic agent for bone disease, the denatonium compound can effectively suppress osteoclast differentiation, and effectively improve osteoporosis by treatment of the denatonium compound in an animal model inducing osteoporosis. And upon confirming that it can be treated, the present invention was completed by first identifying that the denatonium compound can be used as a therapeutic agent for bone disease.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of bone disease, comprising a denatonium (Denatonium) compound or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a health functional food for preventing or improving bone disease, including a denatonium (Denatonium) compound or a salt thereof as an active ingredient.

In order to achieve the object of the present invention as described above, the present invention provides a pharmaceutical composition for the prevention or treatment of bone disease, comprising a denatonium (Denatonium) compound or a pharmaceutically acceptable salt thereof as an active ingredient.

In one embodiment of the present invention, the composition may be to inhibit the formation or differentiation of osteoclasts.

In one embodiment of the present invention, the denatonium (Denatonium) compound or a pharmaceutically acceptable salt thereof, (i) alleviate the loss of spongy bone; (ii) Bone density (BMD), cancellous bone and sebaceous bone volume (BV), volume ratio between cancellous bone and sebaceous bone (BV/TV), cancellous bone thickness (Tb.Th), cancellous bone number (Tb.N), cancellous bone volume (Tb. V), improved reduction of cortical bone volume (Ct.V) and sebaceous bone density (Ct.BMD); Or (iii) improving inhibition of vertebral bone formation; It may have activity.

In one embodiment of the present invention, the bone disease is osteoporosis, facet bone disease, rickets, osteomalacia, renal osteotrophy in patients with renal failure, rheumatoid bone disease, degenerative bone disease, bone metastatic lesion, primary As a result, it may be selected from the group consisting of tumors generated in bone, periodontal disease, inflammatory alveolar bone absorption disease and inflammatory bone absorption disease.

In addition, the present invention provides a dietary supplement for preventing or improving bone disease, which includes a denatonium compound or a salt thereof as an active ingredient.

In one embodiment of the present invention, the denatonium (Denatonium) compound or a salt thereof may be to inhibit the formation or differentiation of osteoclasts.

In one embodiment of the present invention, the denatonium (Denatonium) compound or a salt thereof, (i) alleviate the loss of cancellous bone; (ii) Bone density (BMD), cancellous bone and sebaceous bone volume (BV), volume ratio between cancellous bone and sebaceous bone (BV/TV), cancellous bone thickness (Tb.Th), cancellous bone number (Tb.N), cancellous bone volume (Tb. V), improved reduction of cortical bone volume (Ct.V) and sebaceous bone density (Ct.BMD); Or (iii) improving inhibition of vertebral bone formation; It may have activity.

In addition, in one embodiment of the present invention, the bone disease is osteoporosis, facet bone disease, rickets, osteomalacia, renal osteotrophy in patients with renal failure, rheumatoid bone disease, degenerative bone disease, bone metastatic lesion, It may be primarily selected from the group consisting of tumors generated in bone, periodontal disease, inflammatory alveolar bone absorption disease and inflammatory bone absorption disease.

Denatonium or a salt thereof according to the present invention has excellent activity of inhibiting formation or differentiation of excessive osteoclasts, and (i) spongy bone loss that occurs when inducing bone disease; (ii) Bone density (BMD), cancellous bone and sebaceous bone volume (BV), volume ratio between cancellous bone and sebaceous bone (BV/TV), cancellous bone thickness (Tb.Th), cancellous bone number (Tb.N), cancellous bone volume (Tb. V), reduction in cortical bone volume (Ct.V) and sebaceous bone density (Ct.BMD); (iii) Spine bone formation suppression symptoms can be effectively improved, and thus can be effectively used as a material for medicines and health functional foods that can prevent, improve and treat bone diseases.

Figure 1 is an analysis of the effect on the differentiation of osteoclasts for the denatonium compound of the present invention, inducing osteoclast differentiation by treating RANKL on mouse bone marrow cell-derived macrophages, and the concentration of the denatonium compound of the present invention After the treatment, the number of differentiated osteoclasts was confirmed by TRAP staining.
Figure 2 was administered to each osteoporosis-induced mouse animal model of the present invention by concentration (1.5mg / kg, 4.5mg / kg) by concentration, and then analyzed the distribution of spongy bone through 3D image analysis ( Top left figure), Bone density (BMD), cancellous bone and sebaceous bone volume (BV), cancellous bone to sebaceous volume ratio (BV/TV), cancellous bone thickness (Tb.Th), cancellous bone number (Tb.N) , Spongy bone volume (Tb.V), cortical bone volume (Ct.V), and sebaceous bone density (Ct.BMD) were observed to decrease compared to control mice. In addition, it shows the results of analyzing the spacing between spongy bones (Tb.Sp).
Figure 3 shows the results of analyzing the number of vertebrae by staining method using Alizarin Red S solution after treating the denatonium compound of the present invention for the osteoporosis-induced zebrafish cheers induced by Prednisolon by concentration.

The present invention is characterized by providing a new use of a denatonium compound as a therapeutic agent for bone disease.

Specifically, the present invention can provide a pharmaceutical composition for the prevention or treatment of bone disease, comprising a denatonium (Denatonium) compound or a pharmaceutically acceptable salt thereof as an active ingredient.

The present inventors, while researching to discover new therapeutic agents for bone diseases, confirmed that the denatonium compounds can effectively prevent, improve and treat bone diseases.

According to one embodiment of the present invention, when the macrophages derived from bone marrow cells of a mouse are treated with RANKL and differentiated into osteoclasts, the denatonium compounds of the present invention are treated with osteoclasts. It was confirmed that differentiation was effectively suppressed (see FIG. 1 ).

In another embodiment, when the denatonium compound of the present invention is administered to an actual osteoporosis-induced mouse, bone disease, particularly symptoms of osteoporosis (i) loss of spongy bone; (ii) Bone density (BMD), cancellous bone and sebaceous bone volume (BV), volume ratio between cancellous bone and sebaceous bone (BV/TV), cancellous bone thickness (Tb.Th), cancellous bone number (Tb.N), cancellous bone volume (Tb. V), cortical bone volume (Ct.V) and sebaceous bone bone density (Ct.BMD), it was confirmed that all the symptoms of reduction and suppression (see Fig. 2).

In addition, according to another embodiment, as a result of administering the denatonium (Denatonium) compound of the present invention to the zebra fish cheer model in which osteoporosis is induced, it was found that the number of vertebrae decreased by osteoporosis increases again (Fig. 3). Reference).

Therefore, through these results, the present inventors found that the denatonium (Denatonium) compound can effectively improve, treat and prevent bone disease.

The active ingredient contained in the pharmaceutical composition for the prevention or treatment of bone disease according to the present invention is a denatonium (Denatonium) compound specifically benzyl-[2-(2,6-dimethylanilino)-2-oxoethyl]-diethylazanium Is named.

In addition, the denatonium (Denatonium) compound according to the present invention can be used in the form of a salt, preferably a pharmaceutically acceptable salt, the salt is an acid addition salt formed by a pharmaceutically acceptable free acid (free acid) This is useful.

The expression pharmaceutically acceptable salt is a concentration having a relatively non-toxic and harmless effect on the patient, and any side effect of the base compound of denatonium does not degrade the beneficial effect of the base compound of denatonium due to side effects caused by the salt. It means inorganic addition salt. As the free acid, inorganic and organic acids can be used. As the inorganic acid, hydrochloric acid, bromic acid, nitric acid, sulfuric acid, perchloric acid, and phosphoric acid can be used. As the organic acid, citric acid, acetic acid, lactic acid, maleic acid, and fumarine can be used. Acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, tartaric acid, galacturonic acid, embonic acid, glutamic acid, aspartic acid, oxalic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesul Folic acid, 4-toluenesulfonic acid, salicylic acid, citric acid, benzoic acid or malonic acid can be used. In addition, these salts include alkali metal salts (sodium salt, potassium salt, etc.) and alkaline earth metal salts (calcium salt, magnesium salt, etc.). For example, acid addition salts include acetate, aspartate, benzate, besylate, bicarbonate/carbonate, bisulfate/sulfate, borate, camsylate, citrate, edisylate, esylate, formate, fumarate, Glucetate, Gluconate, Glucuronate, Hexafluorophosphate, Hybenate, Hydrochloride/Chloride, Hydrobromide/Bromide, Hydroiodide/Iodine, Isethionate, Lactate, Malate, Mali Eight, malonate, mesylate, methyl sulfate, naphthylate, 2-naphthylate, nicotinate, nitrate, orotate, oxalate, palmitate, famoate, phosphate/hydrogen phosphate/dihydrogen phosphate, saccharate Late, stearate, succinate, tartrate, tosylate, trifluoroacetate, aluminum, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, Potassium, sodium, tromethamine, zinc salt, etc. may be included.

The acid addition salt according to the present invention is a precipitate produced by dissolving a conventional method, for example, the denatonium compound of the present invention in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, etc. Or by drying under reduced pressure after distilling the solvent and excess acid under reduced pressure, or by crystallization under an organic solvent.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess of an alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the inexpensive compound salt, and evaporating and drying the filtrate. At this time, it is suitable to manufacture sodium, potassium or calcium salts as metal salts. Further, the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (eg, silver nitrate).

Furthermore, the present invention may include all of the denatonium compounds and pharmaceutically acceptable salts thereof, as well as possible solvates, hydrates, isomers, optical isomers, and the like, which can be prepared therefrom.

In addition, the denatonium compound according to the present invention can be isolated from nature or prepared by chemical synthesis methods known in the art.

The pharmaceutical composition for preventing or treating bone disease according to the present invention is a pharmaceutical composition capable of treating bone disease, and may further include a pharmaceutically acceptable carrier. As used herein, “pharmaceutically acceptable” refers to a composition that is physiologically acceptable and does not cause an allergic reaction or similar reaction, such as gastrointestinal disorder, dizziness, etc., when administered to a human. Pharmaceutically acceptable carriers include, for example, oral carriers such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid and the like, and carriers for parenteral administration such as water, suitable oils, saline, aqueous glucose and glycols, etc. Etc. and may further include stabilizers and preservatives. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-parabens and chlorobutanol. As other pharmaceutically acceptable carriers, reference may be made to those described in the following documents (Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995).

The pharmaceutical composition according to the present invention may be formulated in a suitable form according to a method known in the art together with a pharmaceutically acceptable carrier as described above. That is, the pharmaceutical composition of the present invention may be prepared in various parenteral or oral dosage forms according to a known method, and as an example of a parenteral dosage form, an isotonic aqueous solution or suspension is preferred as an injectable dosage form. Injectable formulations can be prepared according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. For example, each component can be formulated for injection by dissolving it in saline or buffer. In addition, formulations for oral administration include, but are not limited to, powders, granules, tablets, pills and capsules.

The pharmaceutical composition formulated in the above manner may be administered through various routes including oral, transdermal, subcutaneous, intravenous or intramuscularly in an effective amount. Means and the route of administration of the substance can be administered through any general route as long as it can reach the target tissue.

In addition, the effective amount in the above refers to an amount that exhibits a preventive or therapeutic effect when administered to a patient. The dosage of the pharmaceutical composition according to the present invention may vary depending on various factors such as the patient's disease type and severity, age, sex, weight, sensitivity to drugs, current treatment type, administration method, target cells, etc. It can be easily determined by experts. In addition, the pharmaceutical composition of the present invention can be administered in combination with a conventional therapeutic agent, can be administered sequentially or simultaneously with a conventional therapeutic agent, and can be administered single or multiple. Preferably, in consideration of all of the above factors, an amount capable of obtaining the maximum effect in a minimal amount without side effects may be administered, more preferably 1-10000 μg/kg body weight/day, even more preferably 10-1000 mg It can be administered repeatedly several times a day with an effective dose of /kg body weight /day.

In the present invention, the bone disease to be prevented, improved, treated, or diagnosed refers to a disease caused by hyperdifferentiation of osteoclasts to cause a disorder of bone tissue or to destroy bone tissue, but the bone disease is not limited thereto. , Osteoporosis, facet bone disease, rickets, osteomalacia, renal osteotrophy in patients with renal failure, rheumatoid bone disease, degenerative bone disease, bone metastatic lesions, primary bone-generated tumors, periodontal disease, inflammatory alveolar bone Absorption disease and inflammatory bone absorption disease.

In addition, since the denatonium compound of the present invention can be added to food for the purpose of preventing and improving symptoms of bone disease, the composition comprising the denatonium compound of the present invention or a salt thereof as an active ingredient may prevent or improve bone disease. It can be used as a food composition for.

Therefore, the food composition for the prevention and improvement of bone diseases containing the denatonium compound of the present invention or a salt thereof as an active ingredient is effective in the prevention and improvement of symptoms of bone disease, for example, the main raw material of food, auxiliary ingredients, food It can be easily used as an additive, functional food or beverage.

As used herein, the term “food” refers to a natural product or a processed product that contains one or more nutrients, and preferably means a state that can be directly eaten through a certain processing process, and has a general meaning. It includes food, food additives, functional food and beverages.

Foods that can be added to the composition for preventing and improving bone disease symptoms according to the present invention include, for example, various foods, beverages, gums, teas, vitamin complexes, and functional foods. In addition, in the present invention, foods include special nutritional foods (e.g., formulas, infants, infant foods, etc.), processed meat products, fish products, tofu, jelly, noodles (e.g. ramen, noodles, etc.), breads, health supplements, seasonings Food (e.g., soy sauce, miso, red pepper paste, mixed sauce, etc.), sauces, confectionery (e.g., snacks), candy, chocolates, gums, ice cream, dairy products (e.g. fermented milk, cheese, etc.), other processed foods, kimchi, Pickled foods (various kimchi, pickles, etc.), beverages (eg, fruit drinks, vegetable drinks, soy milk, fermented beverages, etc.), natural seasonings (eg, ramen soup, etc.) are not limited thereto. The food, beverage or food additive may be prepared by a conventional manufacturing method.

In addition, the "functional food" refers to the food group or food composition that has added value to act and express the function of the food for a specific purpose by using physical, biochemical, and biotechnological techniques, and control the biological defense rhythm of the food composition. Means a food that has been designed and processed to sufficiently express the body control functions related to and recovery, and may be a health functional food. The functional food may include a food-acceptable food supplement additive, and may further include suitable carriers, excipients, and diluents commonly used in the production of functional foods.

In addition, in the present invention, the term "beverage" means a generic term for drinking to relieve thirst or to enjoy a taste, and includes a functional beverage. The beverage is an essential component in the indicated ratio, and there is no particular restriction on other components other than the composition for the prevention and improvement of the symptoms of bone disease, and contains various flavors or natural carbohydrates, etc. can do.

Furthermore, foods containing a composition for the prevention and improvement of bone disease symptoms of the present invention in addition to the above-described flavors, colorants and fillers such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors ( Cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohol, carbonic acid used in carbonated beverages, etc. The components can be used independently or in combination.

In the food containing the composition for preventing and improving bone disease symptoms of the present invention, the amount of the composition according to the present invention may include 0.001% to 90% by weight of the total food weight, preferably 0.1% by weight It may contain from 40 to 40% by weight, and in the case of beverages, it may contain 0.001 g to 2 g, preferably 0.01 g to 0.1 g, based on 100 ml, but for health and hygiene purposes or for health control purposes. In the case of long-term intake to be, it may be below the above range, and the active ingredient is not limited to the above range since it can be used in an amount above the above range because there is no problem in terms of safety.

Hereinafter, the present invention will be described in more detail through examples. These examples are intended to illustrate the present invention more specifically, but the scope of the present invention is not limited to these examples.

<Example 1>

Analysis of the effect of denatonium compounds on osteoclast differentiation

Male 6-week-old C57BL/6 mice were euthanized using 100% CO ₂ gas, and then the femur and tibia were separated. After cutting both ends of the separated tibia and femur, the cells in the bone marrow cavity were collected with a 1 mL syringe, and the red blood cells were dissolved by lysis using a red blood cell lysis buffer (RBCL). The obtained bone marrow cells were then cultured for 3 days in α-MEM medium containing 10% FBS, M-CSF (30 ng/mL), and after 3 days of culture, the attached cells were macrophages (bone marrow-derived macrophages, BMMs). ). After dispensing BMMs cells into a 48-well culture plate, RANKL (100 ng/ml) was added to induce differentiation into osteoclasts. In this process, the denatonium compound of the present invention was concentration-dependent (0.1 mM, 0.2 mM, 0.4). mM) and treated with TRAP staining after 3 days.

As a result, as shown in Figure 1, in the group treated with RANKL alone, BMMs cells showed a significant number of cells differentiated into osteoclasts, whereas in the group treated with the denatonium compound of the present invention, compared with the group treated with RANKL alone It was found that the number of cells differentiated into cells was significantly reduced, and when the 0.2 mM denatonium compound was treated, the osteoclast differentiation was suppressed by 50% or more. In the group treated with a concentration of 0.4 mM, osteoclasts were treated. It has been shown that furnace differentiation is completely suppressed.

Therefore, through these results, the present inventors found that the denatonium compound of the present invention has an activity of effectively inhibiting osteoclast differentiation even in a very small amount.

<Example 2>

Osteoporosis-induced mouse model of the present invention Denatonium Analysis of treatment efficacy of osteoporosis according to compound treatment

The following experiments were performed to analyze osteoporosis prevention or treatment efficacy for the denatonium compound of the present invention. First, the mice used in the experiment were purchased and used for 7-week-old mice at an aged animal growth facility. After stabilization for a week, 8-week-old female mice were randomly separated into 4 groups (4 animals/group), and then experimented. The animals were kept for a week for stable adaptation. Subsequently, 4 mice were configured as a control group, and the rest of the groups were subjected to ovarian resection (OVX), and the experimental animals of the OVX group were injected with 40 mg/kg pentobarbital sodium intraperitoneally and then subjected to general anesthesia to expose both ovaries. And removed to prepare osteoporosis-induced mice. In the normal control group (Sham group), the ovaries were exposed, but not removed. Two weeks after the surgery, the normal control experimental animals ingested the vehicle (water) once a week for 6 weeks, and the experimental group was treated with denatonium in the amount of 1.5 mg/kg body weight and 4.5 mg/kg body weight, respectively. Intraperitoneal injection.

After the administration of the denatonium compound of the present invention to the mice having osteoporosis-induced concentration, it was confirmed whether or not the loss of spongy bone was improved by analyzing the 3D image, and the bone density (BMD), the spongy bone and sebaceous bone volume, which are the analysis indexes of osteoporosis. (BV), the volume ratio between the cancellous bone and sebaceous bone (BV/TV), the thickness of the cancellous bone (Tb.Th), the number of cancellous bones (Tb.N), the cancellous bone volume (Tb.V), cortical bone volume (Ct.V), and the sebaceous bone Bone density (Ct.BMD) of each was analyzed. In addition, Sham mice that did not cause osteoporosis were used as controls.

As a result, as shown in FIG. 2, the 3D image of the femur in the control group (Sham mouse) showed that spongy bone was normally distributed, and that the spongy bone was lost in the OVX mouse (osteoporosis-induced mouse). I could observe.

On the other hand, in the group administered with denatonium to the osteoporosis-induced mice, the low-density denatonium (1.5mg/kg) treated group was found to have a lessening of the cancellous bone loss, and the high-density denatonium (4.5mg/kg) treated group than the low-concentration treated group. It was found that the relief of spongy bone was improved more.

In addition, as a result of analyzing bone indexes through microstructure analysis, in the osteoporosis-induced mouse group, bone density (BMD), spongy bone and sebaceous bone volume (BV), spongy bone and sebaceous bone volume ratio (BV/TV), spongy bone thickness (Tb.Th) ), cancellous bone number (Tb.N), cancellous bone volume (Tb.V), cortical bone volume (Ct.V), and sebaceous bone density (Ct.BMD) were all decreased compared to normal control mice. The spacing between spongy bones (Tb.Sp) was also found to be wider than that of normal control mice.

As a result of treating the osteoporosis-inducing mouse with the denatonium compound of the present invention, the bone density (BMD), an indicator of reduced bone microstructure in the osteoporosis-inducing mouse in both the low (1.5mg/kg) and high (4.5mg/kg) treated groups ), cancellous bone and sebaceous bone volume (BV), cancellous bone and sebaceous bone volume ratio (BV/TV), cancellous bone thickness (Tb.Th), cancellous bone number (Tb.N), cancellous bone volume (Tb.V), cortical bone volume ( Both Ct.V) and sebaceous bone density (Ct.BMD) were found to be significantly recovered, and the interval between cancellous bones (Tb.Sp) was also maintained to be improved and maintained at normal control levels.

Therefore, through these results, the present inventors found that the denatonium compound of the present invention can effectively treat and prevent bone diseases through in vivo animal experiments.

<Example 3>

Osteoporosis-induced Zebra Fish Of the present invention in a cheer model Denatonium For compounds Vertebra Analysis of the impact on formation

The present inventors verified the possibility of using the denatonium compound as a therapeutic agent for bone disease in another animal model. To this end, a 10-day-old fertilized zebrafish pom was treated with 25 μM prednisolone to induce osteoporosis to prepare an osteoporosis-induced zebrafish pom.

After treatment of the osteoporosis-induced zebrafish cheer with the denatonium compound of the present invention by concentration (0.1 mM, 0.2 M, 0.4 mM), fixed with formalin after 3 days, stained with Alizarin Red S solution, and then the number of vertebrae Was measured.

As a result, as shown in FIG. 3, the number of spinal bones was decreased in the zebrafish cheerlead induced by osteoporosis by prednisolon administration, but the number of reduced spinal bones increased in the group treated with the denatonium compound of the present invention. It has been shown to induce the formation of vertebral bones.

Therefore, the denatonium compound of the present invention can actually induce the formation of vertebrae through animal experiments, can suppress the differentiation of osteoclasts, and can restore all bone microstructure analysis indicators to a normal state. As a result, it was found that the denatonium compound can be usefully used as a material for the prevention or treatment of bone diseases.

So far, the present invention has been focused on the preferred embodiments. Those skilled in the art to which the present invention pertains will understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered in terms of explanation, not limitation. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the equivalent range should be interpreted as being included in the present invention.

Claims (8)

Denatonium (Denatonium) compound or a pharmaceutical composition for the prevention or treatment of osteoporosis comprising a pharmaceutically acceptable salt thereof as an active ingredient. According to claim 1,
The composition is characterized in that to inhibit the formation or differentiation of osteoclasts, a pharmaceutical composition for the prevention or treatment of osteoporosis. According to claim 1,
The denatonium (Denatonium) compound or a pharmaceutically acceptable salt thereof,
(i) alleviating the loss of cancellous bone;
(ii) Bone density (BMD), cancellous bone and sebaceous bone volume (BV), volume ratio between cancellous bone and sebaceous bone (BV/TV), cancellous bone thickness (Tb.Th), cancellous bone number (Tb.N), cancellous bone volume (Tb. V), improved reduction of cortical bone volume (Ct.V) and sebaceous bone density (Ct.BMD); or
(iii) improved bone bone formation inhibition; Pharmaceutical composition for the prevention or treatment of osteoporosis, characterized by having an activity. delete Denatonium (Denatonium) compound or a salt thereof as an active ingredient, for preventing or improving osteoporosis health functional food. The method of claim 5,
The denatonium (Denatonium) compound or a salt thereof is characterized in that to inhibit the formation or differentiation of osteoclasts, health functional food for preventing or improving osteoporosis. The method of claim 5,
The denatonium (Denatonium) compound or a salt thereof,
(i) alleviating the loss of cancellous bone;
(ii) Bone density (BMD), cancellous bone and sebaceous bone volume (BV), volume ratio between cancellous bone and sebaceous bone (BV/TV), cancellous bone thickness (Tb.Th), cancellous bone number (Tb.N), cancellous bone volume (Tb. V), improved reduction of cortical bone volume (Ct.V) and sebaceous bone density (Ct.BMD); or
(iii) improved bone bone formation inhibition; Health functional food for preventing or improving osteoporosis, characterized by having activity. delete

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