Novel hydroxyacylnornicotine derivative and uses thereof

Abstract

The present application relates to a novel hydroxyacylnornicotine derivative and uses thereof. A hydroxyacylnornicotine derivative according to one aspect exhibits therapeutic or ameliorative activity for bone diseases by inhibiting differentiation into osteoclasts. Accordingly, the present application provides a novel hydroxyacylnornicotine derivative, a pharmaceutical composition for preventing or treating bone diseases comprising the derivative as an active ingredient, and a health functional food composition for preventing or ameliorating bone diseases.

Description

Novel Hydroxyacylnornicotine Derivatives and Uses thereof

[0001] The present application relates to a novel hydroxyacylnornicotine derivative and the medicinal use of said derivative for the treatment of bone diseases.

[0002] Bone is maintained through a balance between bone formation by osteoblasts and bone resorption by osteoclasts. Maintaining this balance between osteoblasts and osteoclasts is an essential element for maintaining bone homeostasis. The disruption of this balance due to aging occurs when the bone-destroying capacity of osteoclasts exceeds the bone-forming capacity of osteoblasts, thereby breaking down homeostasis. This implies that inhibiting bone resorption by osteoclasts is crucial for the prevention and treatment of age-related bone diseases. Excessive osteoclast activity that outweighs osteoblast activity causes various bone diseases and leads to a decrease in bone mass and structural deterioration of the skeleton.

[0003] Osteoporosis is a skeletal disease characterized by a weakened bone strength due to a decrease in bone mass and changes in microstructure, resulting in a condition prone to fractures. As a skeletal disease caused by various factors, osteoporosis leads to a reduction in bone mass and causes microstructural deformation. However, when estrogen secretion decreases due to menopause, menopausal osteoporosis occurs due to an imbalance between osteoclasts and osteoblasts. Therefore, inhibiting the differentiation of osteoclasts can be an important alternative for treating osteoporosis. Osteoclasts are generated from macrophages derived from hematopoietic stem cells, undergoing various differentiation stages mediated by M-CSF (macrophage colony-stimulating factor) and RANKL (receptor activator of nuclear factor-κB ligand). When RANKL binds to RANK (receptor activator of nuclear factor-κB), an osteoclast receptor present on the surface of osteoclast precursor cells, it induces the activation of c-Fos. Activated c-Fos induces the expression of another transcription factor, NFATc1 (nuclear factor of activated T cell c1). Subsequently, NFATc1 regulates bone resorption, activation, survival, and differentiation by activating the differentiation and function of osteoclasts through the promotion of the expression of transcription factors / osteoclast markers essential for osteoclast differentiation, such as TRAP (tartrate-resistant acid phosphatase).

[0004] Bisphosphonates are currently the most widely used treatments for osteoporosis. However, as numerous side effects have been observed in patients treated with bisphosphonates, concerns regarding this treatment have been raised; therefore, there is a need to develop new treatments that can demonstrate effective therapeutic efficacy with fewer side effects.

[0005] As a result of diligent efforts to develop a therapeutic agent derived from natural products, the inventors isolated a novel hydroxyacylnornicotine derivative from the plant *Nicotiana debenei*. Along with this, they confirmed that the isolated hydroxyacylnornicotine derivative not only exhibits low cytotoxicity but also effectively inhibits differentiation into osteoclasts, thereby being applicable as an active ingredient for the treatment of bone diseases, and based on this, they completed the present invention.

[0006] One aspect is to provide a hydroxyacylnornicotine derivative having an inhibitory effect on differentiation into osteoclasts.

[0007] Another aspect is to provide a pharmaceutical composition for the prevention or treatment of bone diseases comprising the above-mentioned hydroxyacylnornicotine derivative as an active ingredient.

[0008] Another aspect is to provide a health functional food composition for the prevention or improvement of bone diseases comprising the above-mentioned hydroxyacylnornicotine derivative as an active ingredient.

[0009] Another aspect is to provide the use of the hydroxyacylnornicotine derivative for the manufacture of medicines for the prevention or treatment of bone diseases.

[0010] Another aspect is to provide the use of the hydroxyacylnornicotine derivative for the preparation of a health functional food composition for the prevention or improvement of bone diseases.

[0011] Another aspect is to provide a method for preventing, improving, or treating bone disease comprising the step of administering a composition containing the above-mentioned hydroxyacylnornicotine derivative as an active ingredient to an individual in need thereof.

[0012]

[0013] Other objects and advantages of this application will become more apparent from the following detailed description, together with the appended claims and drawings. Anything not described in this specification is omitted, as it can be sufficiently recognized and inferred by those skilled in the art of this application or a similar art field.

[0014] To achieve the above-mentioned purpose of the present application, one aspect provides a compound of Formula I, a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof:

[0015] [Chemical Formula I]

[0016]

[0017] In the above chemical formula I, R1, R3 and R5 are independently hydrogen or hydroxyl groups; and R2 and R4 may independently be hydrogen or C1 to C3 alkyl groups.

[0018] In one embodiment, the compound may be selected from the group consisting of the following compounds: 3,10-dihydroxy-10-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecan-1-one; 3,11-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecan-1-one; 3,10-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecan-1-one; and 3,12-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecan-1-one.

[0019] Another aspect provides a pharmaceutical composition for the prevention or treatment of bone disease comprising, as an active ingredient, a compound of Formula I, a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof.

[0020] Another aspect provides a health functional food composition for the prevention or improvement of bone disease comprising a compound of the above-mentioned formula I, a solvate, a stereoisomer, or a food-grade acceptable salt thereof as an active ingredient.

[0021] In one embodiment, the composition may inhibit differentiation into osteoclasts.

[0022] In one embodiment, the bone disease may be selected from the group consisting of osteoporosis, osteoporotic fracture, osteopenia, osteolytic metastasis, senile kyphosis, Paget's disease, bone defect, nonunion fracture, osteomalacia, osteomalacia fracture, bone formation disorder, osteoarthritis, fibrous osteitis, aplastic bone disease, osteogenesis imperfecta, bone atrophy, periodontal disease, rheumatoid arthritis, metabolic bone disease, and bone damage caused by bone metastasis of cancer cells.

[0023] Another aspect provides the use of a compound of Formula I, a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof for the manufacture of a medicine for the prevention or treatment of bone diseases.

[0024] Another aspect provides the use of a compound of Formula I, a solvate, a stereoisomer, or a food-grade acceptable salt thereof for the preparation of a health functional food composition for the prevention or improvement of bone disease.

[0025] Another aspect provides a method for preventing or treating bone disease comprising the step of administering a composition containing a compound of Formula I, a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof as an active ingredient to an individual in need thereof.

[0026] Another aspect provides a method for preventing or improving bone disease comprising the step of administering a composition containing a compound of Formula I, a solvate, a stereoisomer, or a food-grade acceptable salt thereof as an active ingredient to an individual in need thereof.

[0027] Compounds, solvates, stereoisomers, or pharmaceutically acceptable salts of Formula I according to one aspect may exhibit low cytotoxicity and simultaneously have an inhibitory effect on differentiation into osteoclasts.

[0028] Accordingly, compounds, solvates, stereoisomers, or pharmaceutically acceptable salts of Formula I according to one aspect may be utilized as active ingredients in compositions for improving or treating various bone diseases, including osteoporosis.

[0029] Figure 1 is the result of TLC analysis performed on an ethyl acetate fraction according to one embodiment.

[0030] FIG. 2 is a diagram showing the process of separating and purifying active compounds W21-EA323 and W21-EA324 from an ethyl acetate fraction according to one embodiment.

[0031] Figure 3 is the result of TLC analysis performed on the Sephadex LH-20 column chromatography fraction of ethyl acetate fraction EA-3 according to one embodiment.

[0032] Figure 4 is the result of HPLC analysis performed on the Sephadex LH-20 column chromatography fraction of ethyl acetate fraction EA-3 according to one embodiment.

[0033] Figure 5 is the result of performing a preparative HPLC analysis on the ethyl acetate fraction EA3-2 according to one embodiment.

[0034] Figure 6 is the result of quantifying the active compounds W21-EA323 and W21-EA324 according to one embodiment through HPLC analysis.

[0035] Figure 7 shows the results of quantifying the active compounds W21-EA323 and W21-EA324 according to one embodiment through UV spectrum analysis.

[0036] Figure 8 is the mass spectrum result for the active compound W21-EA323 according to one embodiment.

[0037] FIG. 9 is for an active compound W21-EA323 according to one embodiment 1 This is the H NMR result.

[0038] FIG. 10 is for an active compound W21-EA323 according to one embodiment 13 This is the C NMR result.

[0039] FIG. 11a is for an active compound W21-EA323 according to one embodiment 1 H- 1 This is the H COSY spectrum result.

[0040] FIG. 11b is for an active compound W21-EA323 according to one embodiment 1 H- 1 This is a diagram showing the partial structures (bold lines) within the compound identified through the interpretation of the H COSY spectrum results.

[0041] Figure 12 is the HMQC spectrum result for the active compound W21-EA323 according to one embodiment.

[0042] Figure 13 is the HMBC spectrum result for the active compound W21-EA323 according to one embodiment.

[0043] FIG. 14 shows the chemical structure of the active compound W21-EA323 according to one embodiment. 1 H NMR and 13 This is a result identified through the interpretation of C NMR results.

[0044] Figure 15 shows the results of determining the chemical structure of the active compound W21-EA323 according to one embodiment through the analysis of the two-dimensional NMR spectrum results.

[0045] Figure 16 is the mass spectrum result for the active compound W21-EA324 according to one embodiment.

[0046] FIG. 17 is for an active compound W21-EA324 according to one embodiment 1 This is the H NMR result.

[0047] FIG. 18 is for an active compound W21-EA324 according to one embodiment 13 This is the C NMR result.

[0048] FIG. 19a is for an active compound W21-EA324 according to one embodiment 1 H- 1 This is the H COSY spectrum result.

[0049] FIG. 19b is for an active compound W21-EA324 according to one embodiment 1 H- 1 This is a diagram showing the partial structures (bold lines) within the compound identified through the interpretation of the H COSY spectrum results.

[0050] Figure 20 is the HMQC spectrum result for the active compound W21-EA324 according to one embodiment.

[0051] Figure 21 is the HMBC spectrum result for the active compound W21-EA324 according to one embodiment.

[0052] FIG. 22 shows the chemical structure of the active compound W21-EA324 according to one embodiment. 1 H NMR and 13 This is a result identified through the interpretation of C NMR results.

[0053] Figure 23 shows the results of determining the chemical structure of the active compound W21-EA324 according to one embodiment through the analysis of the two-dimensional NMR spectrum results.

[0054] Figure 24 shows the results of evaluating the inhibitory effect on differentiation into osteoclasts and changes in cell viability by treating bone marrow-derived macrophages with various concentrations of compounds KTD-005 or KTD-006.

[0055] Figure 25 shows the results of microscopic examination of TRAP-stained images after treating bone marrow-derived macrophages with 3.75 μg / ml of compounds KTD-005 or KTD-006.

[0056] Figure 26 shows the results of evaluating the inhibitory effect on differentiation into osteoclasts and changes in cell viability by treating bone marrow-derived macrophages with various concentrations of compounds KTD-007 or KTD-008.

[0057] Figure 27 shows the results of microscopic examination of TRAP-stained images after treating bone marrow-derived macrophages with 3.75 μg / ml of compounds KTD-007 or KTD-008.

[0058] Each description and embodiment disclosed in this application may be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in this application fall within the scope of this application. Furthermore, the scope of this application should not be considered limited by the specific descriptions provided below.

[0059] One aspect provides a compound of the following formula I, a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof.

[0060] [Chemical Formula I]

[0061]

[0062] In the above chemical formula I, R1, R3 and R5 are independently hydrogen or hydroxyl groups; and R2 and R4 may independently be hydrogen or C1 to C3 alkyl groups.

[0063] In this specification, the term “alkyl” means a straight-chain or branched hydrocarbon residue that may be substituted or unsubstituted unless otherwise noted. The alkyl group may include, for example, methyl, ethyl, propyl, or all possible isomers thereof without limitation.

[0064] In this specification, a numerical range indicated by the term "to" refers to a range that includes the values ​​described before and after the term "to" as a lower limit and an upper limit, respectively.

[0065] In this specification, the term “solvate” may mean a compound of the present invention or a salt thereof comprising stoichiometric or non-stoichiometric amounts of solvent bound by non-covalent intermolecular forces. Preferred solvents thereof may be volatile, non-toxic, and / or suitable for administration to humans.

[0066] In this specification, the term "stereoisomer" may mean a compound of the present invention or a salt thereof that has the same chemical formula or molecular formula but is optically or stereochemically different, and specifically, may be a diastereomer, enantiomer, geometric isomer, or shape isomer.

[0067] In addition, the above compound may be used in the form of a pharmaceutically acceptable salt derived from an inorganic acid or an organic acid, for example, the salt may be a salt derived from hydrochloric acid, hydrobromide, sulfuric acid, phosphoric acid, nitric acid, acetic acid, glycolic acid, lactic acid, pyruvate, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, mandelic acid, tartaric acid, citric acid, ascorbic acid, palmitic acid, maleic acid, hydroxymaleic acid, benzoic acid, hydroxybenzoic acid, phenylacetic acid, cinnamic acid, salicylic acid, methanesulfonic acid, benzenesulfonic acid, or toluenesulfonic acid.

[0068] A pharmaceutically acceptable salt of the above compound can be prepared by dissolving the compound of Formula I in a water-miscible organic solvent, such as acetone, methanol, ethanol, or acetonitrile, adding an excess amount of organic acid or an aqueous solution of an inorganic acid, and then precipitating or crystallizing. Subsequently, the solvent or excess acid can be evaporated from the mixture and dried to obtain an addition salt, or the precipitated salt can be prepared by suction filtration.

[0069] In one embodiment, in the above formula I, R1 is a hydroxyl group, R2 is methyl, and R3 to R5 are hydrogen; R3 is a hydroxyl group, R4 is methyl, and R 1, R2 and R5 are hydrogen; R1 is a hydroxyl group, R4 is methyl, and R 2, R3 and R5 are hydrogen; or R5 may be a hydroxyl group, R4 may be methyl, and R1 to R3 may be hydrogen.

[0070] In one embodiment, the compound of Formula I may be selected from the group consisting of the following compounds:

[0071] 3,10-dihydroxy-10-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecan-1-one (3,10-dihydroxy-10-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecan-1-one);

[0072] 3,11-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecan-1-one (3,11-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecan-1-one);

[0073] 3,10-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecan-1-one (3,10-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecan-1-one); and

[0074] 3,12-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecan-1-one (3,12-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecan-1-one).

[0075] In one embodiment, the compound of Formula I may be isolated from Nicotiana debneyi, for example, from the ethyl acetate fraction of the methanol extract of Nicotiana debneyi. However, as described above, the compound may not only be derived from natural products but may also be manufactured using chemical transformation / manufacturing processes well known to those skilled in the art of organic / medicinal chemistry.

[0076]

[0077] Another aspect provides a pharmaceutical composition for the prevention or treatment of bone disease comprising, as an active ingredient, a compound represented by the above chemical formula I, a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof.

[0078] In the above pharmaceutical composition, any terms or elements mentioned that are identical to those already mentioned are as described above.

[0079] In this specification, the term "prevention" refers to any act of suppressing or delaying the onset of a disease through the administration of the above composition.

[0080] In this specification, the term “treatment” means any form of treatment that provides effects to an individual suffering from a disease or at risk of developing a disease, including improvement of the individual’s condition (e.g., one or more symptoms), delay of disease progression, delay of symptom onset, or slowing of symptom progression. Accordingly, “treatment” and “prevention” are not intended to mean the cure or complete elimination of symptoms.

[0081] The "bone disease" targeted for prevention or treatment by the above pharmaceutical composition may be a collective term for diseases caused by an imbalance in the bone metabolic process between osteoblasts and osteoclasts. The bone disease may be, for example, a bone disease associated with excessive bone resorption by osteoclasts or excessive differentiation into osteoclasts, or a bone disease in which therapeutic efficacy can be exerted through the inhibition of osteoclast activity or differentiation into osteoclasts. Specifically, the bone disease may be selected from the group consisting of osteoporosis, osteoporotic fracture, osteopenia, osteolytic metastasis, senile kyphosis, Paget's disease, bone defect, nonunion fracture, osteomalacia, osteomalacia fracture, bone formation disorder, osteoarthritis, fibrous osteitis, aplastic bone disease, osteogenesis imperfecta, bone atrophy, periodontal disease, rheumatoid arthritis, metabolic bone disease, and bone damage caused by bone metastasis of cancer cells, but is not limited thereto.

[0082] Among the aforementioned bone diseases, "osteoporosis" refers to a pathological condition in which bone mass decreases and bone strength weakens due to qualitative changes, making fractures highly likely. In the case of osteoporosis, the disease develops and progresses as the balance between the activities of osteoblasts and osteoclasts is disrupted, leading to excessive bone destruction by osteoclasts. A major molecule in osteoporosis is RANKL (receptor-activator of nuclear factor kappa-B ligand), and it is known that RANKL binds to its receptor to activate various transcription factors, thereby promoting the differentiation of osteoclasts. Furthermore, it has been reported that when macrophages are treated with RANKL, they differentiate into TRAP (Tartrate-resistant acid phosphatase)-positive cells along with binding to RANK (Receptor-activator of nuclear factor kappa-B); when these TRAP-positive cells are stimulated with inflammatory factors such as RANKL and TNF-α, the cells fuse together and differentiate into multinucleated TRAP-positive cells. Under this technical background, the TRAP can be applied to evaluate the biological / pharmacological activity of an active compound according to one embodiment as one of the differentiation markers into osteoclasts.

[0083] According to one embodiment, the hydroxyacylnornicotine derivative isolated and purified in one embodiment not only did not exhibit cytotoxicity but was also able to inhibit differentiation into osteoclasts. Therefore, the hydroxyacylnornicotine derivative can be applied to regulate the imbalance in the bone metabolic process between osteoblasts and osteoclasts. Through this, the hydroxyacylnornicotine derivative can be utilized for medicinal purposes for the prevention or treatment of bone diseases, and specifically, can be included as an active ingredient in a pharmaceutical composition for the prevention or treatment of bone diseases.

[0084] In this specification, the term “included as an active ingredient” means that a compound, solvate, stereoisomer, or pharmaceutically acceptable salt thereof represented by Formula I is added to an extent that can produce the effects mentioned in this specification, and includes formulation in various forms by adding various ingredients as auxiliary ingredients for drug delivery and stabilization.

[0085] In one embodiment, the pharmaceutical composition may include a conventionally pharmaceutically acceptable carrier, excipient, or additive. The pharmaceutical composition may be formulated according to conventional methods and may be prepared in various oral administration forms such as tablets, pills, powders, capsules, syrups, emulsions, microemulsions, or parenteral administration forms such as intramuscular, intravenous, or subcutaneous administration.

[0086] When the above pharmaceutical composition is prepared in the form of an oral formulation, examples of additives or carriers used include cellulose, calcium silicate, corn starch, lactose, sucrose, dextrose, calcium phosphate, stearic acid, magnesium stearate, calcium stearate, gelatin, talc, surfactants, suspending agents, emulsifiers, diluents, etc. When the pharmaceutical composition of the present invention is prepared in the form of an injectable formulation, examples of additives or carriers include water, saline solution, glucose aqueous solution, similar sugar aqueous solution, alcohol, glycol, ether (e.g., polyethylene glycol 400), oil, fatty acid, fatty acid ester, glyceride, surfactant, suspending agent, emulsifier, etc.

[0087] The dosage of the above pharmaceutical composition is an amount effective for the treatment or prevention of an individual or patient, and may be administered orally or parenterally as intended. When administered orally, it may be administered in an amount of 0.01 to 1000 mg per kg of body weight per day based on the active ingredient, more specifically 0.1 to 300 mg; when administered parenterally, it may be administered in an amount of 0.01 to 100 mg per kg of body weight per day based on the active ingredient, more specifically 0.1 to 50 mg. The dosage for a specific individual or patient should be determined in light of various relevant factors such as the patient's body weight, age, gender, health status, diet, time of administration, method of administration, and severity of the disease, and it should be understood that it may be appropriately adjusted by a professional. The above dosage is not intended to limit the scope of the present invention in any way. A physician or veterinarian with ordinary skills in the relevant technical field can easily determine and prescribe the required effective amount of the pharmaceutical composition. For example, a physician or veterinarian may start the dosage of the compound of the present invention used in a pharmaceutical composition at a level lower than that required to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved.

[0088] In one embodiment, the pharmaceutical composition includes within its scope a pharmaceutical composition comprising, as an active ingredient, at least one of the compounds according to one embodiment alone or in combination with a pharmaceutical carrier in a therapeutically effective amount. Optionally, the compound according to one embodiment may be administered alone, in combination with a compound according to another embodiment, or simultaneously, separately, or sequentially with one or more other therapeutic agents, e.g., a bone disease treatment or other pharmaceutically active substance.

[0089]

[0090] Another aspect provides a health functional food composition for the prevention or improvement of bone disease, comprising as an active ingredient a compound represented by the above chemical formula I, a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof.

[0091] In the above-mentioned health functional food composition, any terms or elements mentioned that are identical to those already mentioned are as described above.

[0092] In this specification, the term "improvement" refers to any action that at least reduces parameters related to the condition being treated, such as the severity of symptoms. In this case, the health functional food composition may be used for the prevention or improvement of a target disease, either simultaneously with or separately from a therapeutic agent, either before or after the onset of the disease.

[0093] The above-described health functional food composition may be used in combination with a food / ingredient containing the active compound, or with other foods or food ingredients, and may be used appropriately according to conventional methods. The amount of the active ingredient may be appropriately determined according to the purpose of use (prevention, health, or therapeutic treatment). Generally, when manufacturing food or beverages, the composition of the present invention is added in an amount of 15% by weight or less, preferably 10% by weight or less, relative to the raw materials. However, in the case of long-term consumption for the purpose of health and hygiene or health control, the above amount may be less than the above range.

[0094] In addition to containing the active ingredient as an essential component, the above-mentioned health functional food composition has no special restrictions on the type of health functional food or additional ingredients. Among the types of health functional foods, beverage compositions may contain various flavoring agents or natural carbohydrates as additional ingredients, similar to ordinary beverages. The above-mentioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As sweeteners, natural sweeteners such as taumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used. The above health functional food composition may also contain nutritional supplements, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, or combinations thereof. The above health functional food composition may also contain fruit pulp for the production of natural fruit juice, fruit juice beverages, or vegetable beverages, or combinations thereof.

[0095]

[0096] Another aspect provides the use of a compound of Formula I, a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof for the manufacture of a medicine for the prevention or treatment of bone diseases.

[0097] Another aspect provides the use of a compound of Formula I, a solvate, a stereoisomer, or a food-grade acceptable salt thereof for the preparation of a health functional food composition for the prevention or improvement of bone disease.

[0098] In the above use, any terms or elements mentioned that are identical to those already mentioned are as stated above.

[0099]

[0100] Another aspect provides a method for preventing or treating a bone disease comprising the step of administering to an individual in need a compound of Formula I, a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof, or a composition containing the same as an active ingredient.

[0101] Another aspect provides a method for preventing or improving bone disease comprising the step of administering to an individual in need a compound of Formula I, a solvate, a stereoisomer, or a food-grade acceptable salt thereof, or a composition containing the same as an active ingredient.

[0102] In the above method, any terms or elements mentioned that are identical to those already mentioned are as described above.

[0103] In this specification, the term “individual” refers to a subject requiring treatment for a disease, and more specifically, to mammals such as human or non-human primates, mice, dogs, cats, horses, and cattle.

[0104] In this specification, the terms “administering” and “applying” are used interchangeably and may mean causing at least partial localization of the composition according to one embodiment to a desired site, or dispensing the composition according to one embodiment into an individual by a route of administration.

[0105] Administration may be carried out by methods known in the art. Methods of administration, such as the route of administration and frequency of administration, may be appropriately selected by a person skilled in the art. Administration may be administered directly to an individual by any means, for example, via routes such as intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal, or subcutaneous administration. The administration may be systemic or local.

[0106] The dosage may vary depending on factors such as the formulation method, method of administration, patient's age, body weight, gender, pathological condition, food, time of administration, route of administration, and response sensitivity, and a person skilled in the art can appropriately adjust the dosage by considering these factors. The frequency of administration may be once a day or two or more times within the range of clinically acceptable side effects, and the administration site may be one or two or more sites. The total duration of administration may range from 1 to 30 days per treatment, administered daily or at intervals of 2 to 5 days. If necessary, the same treatment may be repeated after the appropriate time. For animals other than humans, the dosage may be the same as that for humans per kg, or an amount calculated by converting the above dosage based on, for example, the ratio of organ (e.g., heart) volumes between the target animal and humans (e.g., average value).

[0107]

[0108] The present invention will be explained in more detail below through examples. However, these examples are intended to illustrate the invention and the scope of the invention is not limited to these examples.

[0109]

[0110] Preparation Example 1. Isolation and purification of active ingredients derived from Nicotiana debenei extract

[0111] 1-1. Preparation of Ethyl Acetate Fraction Derived from Nicotiana Devnay Extract

[0112] The ground above-ground parts of Nicotiana debenay were extracted with a methanol solvent, and fractions were obtained for each solvent. Specifically, 900 g of the ground above-ground parts of Nicotiana debenay sample (W21) was added to 9 L of methanol, and a total of 18 L of methanol extract obtained by repeated extraction was concentrated under reduced pressure to obtain a concentrate. An equal amount of hexane was added to the concentrate and partitioned to separate it into a hexane layer and a water layer. Subsequently, ethyl acetate was added to the water layer and partitioned to separate it into an ethyl acetate layer and a water layer. Subsequently, the ethyl acetate layer exhibiting activity was concentrated and silica gel column chromatography was performed (elumination solvent, chloroform-methanol = 50:1, 20:1, 10:1, 5:1, 2:1, 1:1, v / v, eluted in 500 mL each), and as shown in Figure 1, the results were fractionated into 8 groups (EA-1 to EA-8) through thin layer chromatography (TLC) analysis.

[0113]

[0114] 1-2. Isolation and Purification of Active Compounds Derived from Fraction EA-3

[0115] In this preparation example, as shown in Fig. 2, the active compound was isolated and purified from fraction EA-3, which exhibits activity among the ethyl acetate fractions.

[0116] Specifically, the active fraction EA-3 was concentrated and dissolved in DMSO, and ODS-MPLC (eluent solvent, 0→100% aq. methanol gradient) was performed as shown in Fig. 3. Subsequently, as shown in Fig. 4, high-performance liquid chromatography (HPLC) was performed on each eluent to fractionate it into three groups (EA3-1, EA3-2, EA3-3). Afterward, the active fraction EA3-2 was concentrated and dissolved in methanol, followed by preparative ODS-HPLC. As a result, as shown in Fig. 5, two active compounds, W21-EA323 and W21-EA324, were separated. Here, the operating conditions of the above-mentioned preparative reversed-phase HPLC (HPLC system HITACHI EZchrome, HITACHI, Pump L2130, Autosampler L2200, PDA Detector L2455) are as shown in Table 1 below.

[0117] [Table 1]

[0118]

[0119] Subsequently, as shown in FIGS. 6 and 7, HPLC chromatograms and UV spectra were performed. The conditions for performing the HPLC chromatogram (HITACHI Chromaster, HITACHI, Pump 5110, Autosampler 5210, Column Oven 5310, PDA Detector 5430) are as shown in Table 2 below, and as a result, the separated and purified active compounds W21-EA323 and W21-EA324 were quantified.

[0120] [Table 2]

[0121]

[0122]

[0123] Experimental Example 1. Structural analysis of the active compound

[0124] 1-1. Active compound W21-EA323

[0125] (1) Mass spectrum measurement

[0126] As a result of measuring the high-resolution ESI-mass spectrum in positive mode, as shown in Fig. 8, [M+H] at m / z377.2769 + A peak was observed, confirming the molecular weight as 376, and the molecular formula was C 22 H 36 It was identified as N2O3.

[0127] (2) 1 H NMR spectrum measurement

[0128] 1 As a result of measuring the H NMR spectrum, as shown in Fig. 9, the compound was estimated to be a mixture of two conformation isomers, and the ratio 1The integral value of the H NMR peaks was approximately 1.0:0.5. As a result of analyzing the template peaks, aromatic methine protons were observed at 8.49 (2H, br. s), 7.48 (d, J=7.5 Hz), and 7.30 ppm; methine protons bound to nitrogen or oxygen at 5.20 (dd, J=8.0, 3.5 Hz) and 4.00 (m) ppm; methylene protons bound to nitrogen at 3.70 / 3.62 ppm; multiple methylene protons between 1.1 and 2.6 ppm; and two methyl protons at 1.12 and 0.87 ppm. As a result of analyzing the peaks of the subtypes, aromatic methine protons were observed at 8.55 (2H, br. s), 7.45 (d, J=8.0 Hz), and 7.30 ppm; methine protons bound to nitrogen or oxygen at 4.94 (br. d, J=7.5 Hz) and 3.91 (m) ppm; methylene protons bound to nitrogen at 3.73 ppm; multiple methylene protons between 1.1 and 2.5 ppm; and two methyl protons at 1.11 and 0.87 ppm. In addition to these signals, methyl proton peaks originating from other acyl chains were observed at 1.18 and 1.19 ppm.

[0129] (3) 13 C NMR spectrum measurement

[0130] 13 As a result of measuring the 3C NMR spectrum, as shown in Fig. 10, two sets of peaks with similar chemical shift values ​​but different intensities were observed. Analysis of the template peaks revealed a carbonyl carbon at 171.8 ppm and four sp⁻¹ peaks at 148.1, 147.4, 133.4, and 124.2 ppm. 2 methine carbon, one sp at 138.7 ppm 2Carbon, an oxygenated quaternary carbon at 72.9 ppm, an oxygenated methine carbon at 67.8 ppm, nitrogen-bonded carbons at 58.6 and 47.6 ppm, and additionally, multiple methylene carbons between 23 and 42 ppm, and methyl carbons at 26.4 and 8.2 ppm were observed. Analysis of the sub-peaks revealed a carbonyl carbon at 172.7 ppm, and four sp⁻¹s at 149.1, 147.4, 132.9, and 124.2 ppm. 2 methine carbon, one sp at 138.2 ppm 2 Carbon, oxygenated quaternary carbon at 72.8 ppm, oxygenated methine carbon at 67.9 ppm, carbon bonded to nitrogen at 59.4 and 46.9 ppm, and in addition, multiple methylene carbons between 21 and 42 ppm, and methyl carbons at 26.4 and 8.2 ppm were observed. In addition to these signals, oxygenated quaternary carbon at 71.0 ppm, methylene carbon at 43.9 ppm, and methyl carbon at 29.2 ppm, originating from other acyl chains, were observed.

[0131] (4) 1 H- 1 H COSY spectrum measurement

[0132] 1 H- 1 As a result of measuring the H COSY spectrum, the substructures of the main and secondary isomers were determined, as shown in Figures 11a and 11b. From the above results, it was confirmed that the compound is a mixture of hydroxyacylnornicotine (HOAcylNN) in which an acyl chain is attached to the structure of nornicotine.

[0133] (5) Measurement of HMQC and HMBC spectrum

[0134] As a result of measuring the HMQC spectrum, as shown in Fig. 12, the proton-bearing carbon constituting the present compound ( 1 J C-HThe chemical structure of the compound was determined from the long-range correlations observed from each hydrogen as a result of measuring the HMBC spectrum, as shown in Fig. 13. Specifically, in the case of the template form, a nornicotine moiety was revealed through long-range correlations observed from the 7.48 ppm methine proton to carbons at 148.1, 147.4, and 58.6 ppm, and from the 5.20 ppm methine proton to carbons at 147.4 and 133.4 ppm. In addition, a long-range correlation was observed from the 2.52 / 2.35 ppm methylene proton to the 171.8 ppm carbonyl carbon, and from the 1.12 ppm methyl proton to carbons at 72.9, 41.3, and 34.2 ppm, thereby revealing the structure of the acyl chain terminal. In the case of the morphology, a nornicotine moiety was revealed through long-range correlations observed between the 7.45 ppm methine proton and carbons at 149.1, 147.4, and 59.4 ppm, and between the 4.94 ppm methine proton and carbons at 147.4, 138.2, 132.9, and 46.9 ppm. Additionally, long-range correlations were observed between the 2.37 / 1.81 ppm methylene proton and carbonyl carbon at 172.7 ppm, and between the 1.11 ppm methyl proton and carbons at 72.8, 41.3, and 34.2 ppm, thereby elucidating the structure of the acyl chain terminal. Furthermore, fragment peaks 149.1069 and 175 were observed in the high-resolution ESI-mass spectrum.It was confirmed from 0861 that the acyl chain is bonded to nitrogen. Although the proton and carbon peaks of the methylene group constituting the acyl chain were observed to overlap, the structure of the compound was identified as a hydroxyacylnornicotine (HOAcylNN) mixture based on the molecular weight and molecular formula.

[0135] That is, the aforementioned 1 H NMR and 13 The chemical structures identified by the interpretation of the 1 / 2D NMR spectrum and the 2D NMR spectrum are shown in FIGS. 14 and 15. In the following examples, regarding the identified compounds, 3,10-dihydroxy-10-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecane-1-one was named KTD-005 and 3,11-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecane-1-one was named KTD-006.

[0136]

[0137] 1-2. Active compound W21-EA324

[0138] (1) Mass spectrum measurement

[0139] As a result of measuring the high-resolution ESI-mass spectrum in positive mode, as shown in Fig. 16, [M+H] at m / z377.2767 + A peak was observed, confirming the molecular weight as 376, and the molecular formula was C 22 H 36 It was identified as N2O3.

[0140] (2) 1 H NMR spectrum measurement

[0141] 1As a result of measuring the H NMR spectrum, as shown in Figure 17, two sets of peaks with similar chemical shift values ​​but different intensities were observed. Upon analysis of the template peaks, aromatic methine protons were observed at 8.51, 7.48 (d,J=7.5 Hz), and 7.30 ppm; methine protons bound to nitrogen or oxygen at 5.20 (dd,J=8.5, 3.5 Hz), 4.01 (m), and 3.33 ppm; methylene protons bound to nitrogen at 3.70 / 3.61 ppm; multiple methylene protons between 1.1 and 2.6 ppm; and two methyl protons at 0.91 and 0.89 ppm. As a result of analyzing the peaks of the subtypes, aromatic methine protons were observed at 8.51, 7.45 (d,J=7.5 Hz), and 7.30 ppm; methine protons bound to nitrogen or oxygen at 4.95 (br. d,J=7.5 Hz), 3.91 (m), and 3.33 ppm; methylene protons bound to nitrogen at 3.75 ppm; multiple methylene protons between 1.1 and 2.5 ppm; and two methyl protons at 0.91 and 0.89 ppm. In addition to these signals, peaks of methylene protons bound to oxygen at 3.49 / 3.40 ppm and methyl protons at 0.90 ppm, originating from other acyl chains, were observed.

[0142] (3) 13 C NMR spectrum measurement

[0143] 13As a result of measuring the 3C NMR spectrum, as shown in Fig. 18, two sets of peaks with similar chemical shift values ​​but different intensities were observed. Analysis of the template peaks revealed carbonyl carbon at 171.8 ppm, and sp⁻ at 148.3, 147.1, and 133.4 ppm. 2 methine carbon, one sp at 138.3 ppm 2 Carbon, oxygenated methine carbon at 76.8 and 67.8 ppm, nitrogen-bonded carbon at 58.6 and 47.6 ppm, multiple methylene carbons between 23 and 41 ppm, and methyl carbons at 18.9 and 17.1 ppm were observed. Analysis of the sub-morphological peaks revealed a carbonyl carbon at 172.5 ppm and sp at 149.0, 147.4, and 132.9 ppm. 2 Methine carbon, oxygenated methine carbon at 76.7 and 67.9 ppm, carbon combined with nitrogen at 59.4 and 46.9 ppm, and a number of methylene carbons between 21 and 41 ppm, and methyl carbons at 18.9 and 17.1 ppm were observed.

[0144] (4) 1 H- 1 H COSY spectrum measurement

[0145] 1 H- 1 As a result of measuring the H COSY spectrum, the partial structures of the main and secondary isomers were determined, as shown in Figures 19a and 19b. From the above results, it was confirmed that the compound is a hydroxyacylnornicotine (HOAcylNN) mixture in which an acyl chain is attached to the structure of nornicotine.

[0146] (5) Measurement of HMQC and HMBC spectrum

[0147] As a result of measuring the HMQC spectrum, as shown in Fig. 20, the proton-bearing carbon constituting the present compound ( 1 J C-HThe chemical structure of the compound was determined from the long-range correlations observed from each hydrogen as a result of measuring the HMBC spectrum, as shown in Fig. 21. Specifically, in the case of the template form, long-range correlations were observed from the 7.48 ppm methine proton to carbons at 148.3, 147.1, and 58.6 ppm; from the 5.20 ppm methine proton to carbon at 23.7 ppm; and from the 2.34 / 1.88 ppm methylene proton to carbon at 138.3 ppm, revealing a nornicotine moiety. In addition, long-range correlations were observed between the 2.53 / 2.38 ppm methylene proton and the 171.8 ppm carbonyl carbon, and between the 0.89 and 0.91 ppm methyl protons and the 76.8 and 18.9 ppm and 76.8 and 17.1 ppm carbons, respectively, thereby elucidating the structure of the acyl chain terminal. In the case of the subform, long-range correlations were observed between the 7.45 ppm methine proton and the 149.0, 147.4, and 59.4 ppm carbons, and between the 4.95 ppm methine proton and the 147.4, 132.9, 46.9, and 21.6 ppm carbons, thereby revealing a nornicotine moiety. In addition, long-range correlations were observed between the 172.5 ppm carbonyl carbon from the 2.35 / 1.82 ppm methylene proton and between the 76.7 and 18.9 ppm and 76.7 and 17.1 ppm carbons from the 0.89 and 0.91 ppm methyl protons, respectively, thereby elucidating the structure of the acyl chain terminal.In addition, it was determined from fragment peaks 149.1070 and 175.0862 observed in the high-resolution ESI-mass spectrum that the acyl chain is bonded to nitrogen. Although the proton and carbon peaks of the methylene group constituting the acyl chain were observed to overlap, the structure of the compound was identified as a hydroxyacylnornicotine (HOAcylNN) mixture based on its molecular weight and molecular formula.

[0148] That is, the aforementioned 1 H NMR and 13 The chemical structures identified by the interpretation of the 1 / 2D NMR spectrum and the 2D NMR spectrum are shown in FIGS. 22 and 23. In the following examples, regarding the identified compounds, 3,10-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecane-1-one was named KTD-007 and 3,12-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecane-1-one was named KTD-008.

[0149]

[0150] Experimental Example 2. Evaluation of the inhibitory effect on differentiation into osteoclasts

[0151] 2-1. Active compound W21-EA323 (Compounds KTD-005, KTD-006)

[0152] In this experimental example, the effect of compounds KTD-005 or KTD-006 according to one embodiment on osteoclast differentiation was confirmed. To evaluate the inhibitory activity on osteoclast differentiation, bone marrow-derived macrophages obtained by culturing mouse bone marrow cells in α-MEM medium containing M-CSF (50 ng / ml) and 10% FBS for 3 days were used as osteoclast progenitor cells. To generate osteoclasts, the obtained bone marrow-derived macrophages were placed at a rate of 1 × 10⁶ per well in a 96-well plate.4 After aliquoting, the cells were cultured for 3 days in medium containing M-CSF (50 ng / mL) and RANKL (50 ng / mL). Under the same culture conditions, bone marrow-derived macrophages were cultured with the addition of compounds KTD-005 or KTD-006 at various concentrations (0.47, 0.94, 1.88, 3.75, 7.5, 15 μg / mL). Subsequently, the cells were fixed with 10% formalin, and TRAP (Tartrate-resistant acid phosphatase) was stained using naphthol AS-MS phosphate and Fast red violet LB salt. Additionally, to evaluate cell viability and cytotoxicity, bone marrow-derived macrophages were cultured at a density of 1 × 10⁶ per well in a 96-well plate. 4 After dispensing individual cells, the cells were cultured for 2 days in a medium containing M-CSF (50 ng / ml) with compounds KTD-005 or KTD-006 at various concentrations (0.47, 0.94, 1.88, 3.75, 7.5, 15 μg / ml). Cell viability was evaluated by measuring absorbance at 450 nm using Cell Counting Kit-8 (Dojindo Inc.). Meanwhile, in this experiment, the untreated group (Control) was used as the control, and the group treated only with RANKL (RANKL) was used as the positive control.

[0153] As a result, as shown in FIGS. 24 and 25, the group treated with compound KTD-005 or KTD-006 according to one embodiment showed a decrease in TRAP, a differentiation marker for osteoclasts, without a significant decrease in cell viability, and the level of TRAP showed a tendency to decrease depending on the treatment concentration of the active compound.

[0154]

[0155] 2-2. Active compound W21-EA324 (Compounds KTD-007, KTD-008)

[0156] In this experimental example, the effect of compound KTD-007 or KTD-008 according to one embodiment on osteoclast differentiation was confirmed in the same manner as in Experimental Example 2-1 above. Meanwhile, in this experimental example, an untreated group (Control) and a group treated only with RANKL (RANKL) were used as the positive control.

[0157] As a result, as shown in FIGS. 26 and 27, the group treated with compound KTD-007 or KTD-008 according to one embodiment showed a decrease in TRAP, a differentiation marker for osteoclasts, without a significant decrease in cell viability, and the level of TRAP showed a tendency to decrease depending on the treatment concentration of the active compound.

[0158] The foregoing description of the present invention is for illustrative purposes only, and those skilled in the art will understand that other specific forms can be easily modified without altering the technical spirit or essential features of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

Claims

1. Compounds, solvates, stereoisomers, or pharmaceutically acceptable salts of the following chemical formula I: [Chemical Formula I] In the above chemical formula I, R1, R3, and R5 are independently hydrogen or hydroxyl groups; and R2 and R4 are independently hydrogen or C1 to C3 alkyl groups.

2. The compound of the following formula I, solvate, stereoisomer, or pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of the following compounds: 3,10-dihydroxy-10-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecan-1-one; 3,11-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecan-1-one; 3,10-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecan-1-one; and 3,12-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecan-1-one.

3. A pharmaceutical composition for the prevention or treatment of bone diseases comprising, as an active ingredient, a compound of Formula I below, a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof: [Chemical Formula I] In the above chemical formula I, R1, R3, and R5 are independently hydrogen or hydroxyl groups; and R2 and R4 are independently hydrogen or C1 to C3 alkyl groups.

4. A pharmaceutical composition according to claim 3, wherein the compound is selected from the group consisting of the following compounds: 3,10-dihydroxy-10-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecane-1-one; 3,11-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecane-1-one; 3,10-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecane-1-one; and 3,12-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecane-1-one.

5. The pharmaceutical composition of claim 3, wherein the pharmaceutical composition inhibits differentiation into osteoclasts.

6. A pharmaceutical composition according to claim 3, wherein the bone disease is selected from the group consisting of osteoporosis, osteoporotic fracture, osteopenia, osteolytic metastasis, senile kyphosis, Paget's disease, bone defect, nonunion fracture, osteomalacia, osteomalacia fracture, bone formation disorder, osteoarthritis, fibrous osteitis, aplastic bone disease, osteogenesis imperfecta, bone atrophy, periodontal disease, rheumatoid arthritis, metabolic bone disease, and bone damage caused by bone metastasis of cancer cells.

7. A health functional food composition for the prevention or improvement of bone diseases comprising, as an active ingredient, a compound of Chemical Formula I below, a solvate, a stereoisomer, or a food-grade acceptable salt thereof: [Chemical Formula I] In the above chemical formula I, R1, R3, and R5 are independently hydrogen or hydroxyl groups; and R2 and R4 are independently hydrogen or C1 to C3 alkyl groups.

8. A health functional food composition according to Claim 7, wherein the compound is selected from the group consisting of the following compounds: 3,10-dihydroxy-10-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecane-1-one; 3,11-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecane-1-one; 3,10-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecane-1-one; and 3,12-dihydroxy-11-methyl-1-(2-(pyridin-3-yl)pyrrolidin-1-yl)dodecane-1-one.

9. A health functional food composition according to claim 7, wherein the bone disease is selected from the group consisting of osteoporosis, osteoporotic fracture, osteopenia, osteolytic metastasis, senile kyphosis, Paget's disease, bone defect, nonunion fracture, osteomalacia, osteomalacia fracture, bone formation disorder, osteoarthritis, fibrous osteitis, aplastic bone disease, osteogenesis imperfecta, bone atrophy, periodontal disease, rheumatoid arthritis, metabolic bone disease, and bone damage caused by bone metastasis of cancer cells.

10. Use of compounds, solvates, stereoisomers, or pharmaceutically acceptable salts thereof of the following formula I for the manufacture of medicines for the prevention or treatment of bone diseases: [Chemical Formula I] In the above chemical formula I, R1, R3, and R5 are independently hydrogen or hydroxyl groups; and R2 and R4 are independently hydrogen or C1 to C3 alkyl groups.

11. Use of compounds, solvates, stereoisomers, or food-grade acceptable salts thereof of the following formula I for the preparation of a health functional food composition for the prevention or improvement of bone diseases: [Chemical Formula I] In the above chemical formula I, R1, R3, and R5 are independently hydrogen or hydroxyl groups; and R2 and R4 are independently hydrogen or C1 to C3 alkyl groups.

12. A method for preventing or treating a bone disease comprising the step of administering a composition comprising, as an active ingredient, a compound of Formula I below, a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof to an individual in need thereof: [Chemical Formula I] In the above chemical formula I, R1, R3, and R5 are independently hydrogen or hydroxyl groups; and R2 and R4 are independently hydrogen or C1 to C3 alkyl groups.

13. A method for preventing or improving bone disease comprising the step of administering to an individual in need a composition comprising, as an active ingredient, a compound of Formula I below, a solvate, a stereoisomer, or a food-grade acceptable salt thereof: [Chemical Formula I] In the above chemical formula I, R1, R3, and R5 are independently hydrogen or hydroxyl groups; and R2 and R4 are independently hydrogen or C1 to C3 alkyl groups.

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