KR20100130015A - Pharmaceutical composition for the prevention or treatment of osteoporosis or obesity comprising a chloroimidazole derivative or a pharmaceutically acceptable salt thereof as an active ingredient - Google Patents

Abstract

PURPOSE: A pharmaceutical composition containing chloroimidazole derivative for preventing or treating osteoporosis, obesity, diabetes or hyperlipidemia is provided to control TAZ(transcriptional coactivator with PDZbinding motif) protein. CONSTITUTION: A pharmaceutical composition for preventing or treating osteoporosis contains a compound of chemical formula 1 or pharmaceutically acceptable salt thereof as an active ingredient. A pharmaceutical composition for obesity, diabetes or hyperlipidemia contains the compound or pharmaceutically acceptable salt as an active ingredient. The compound is chloroimidazole derivatives.

Description

Pharmaceutical composition for the prevention or treatment of osteoporosis or obesity comprising a chloroimidazole derivative or a soluble acceptable salt know as an active ingredient}

The present invention relates to a pharmaceutical composition for the prevention or treatment of osteoporosis, obesity, diabetes or hyperlipidemia comprising a chloroimidazole derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

As the population ages, senile diseases are on the rise, and in advanced countries with a aging population, it is more serious. In particular, osteoporosis is a representative senile disease, a disease that most elderly people experience. Osteoporosis is a condition in which the amount of bone is reduced and the bone strength is weakened due to qualitative changes, which is likely to cause fractures. This is caused by a breakdown of the balance between bone absorption and bone formation, due to the excessive progression of bone absorption than bone formation. Lime of the bone tissue is reduced, the dense thinning of the bone, thereby widening the bone marrow cavity (骨髓 腔), and as the symptoms progress, the bone weakens, leading to a state of fragile fracture even at a small impact. Bone tissue is a dynamic tissue that is formed by osteoblasts and constantly breaks down and absorbed by osteoclasts.

In the past, the study was mainly focused on bone minerals, namely, calcium and phosphorus metabolic abnormalities, but no significant progress was made in determining the pathogenesis. Materials currently used for the treatment of osteoporosis include bisphosphonate preparations (alendronate, ethidronate), hormone preparations (raloxyphene), vitamin D preparations, calcitonin preparations, calcium preparations and the like. However, the bisphosphonate formulations have a problem of poor absorption, difficult to take, and cause esophagitis, and hormonal formulations have to be taken for a lifetime, and long-term administration may cause side effects such as breast cancer, uterine cancer, gallstones and thrombosis. In addition, vitamin D formulations have the disadvantage of being expensive and indeterminate effects, calcitonin formulations are expensive and difficult to administer, and calcium formulations have fewer side effects but are limited to preventive effects rather than treatment. In addition, osteoporosis cannot be treated with short-term administration of the drug alone, and long-term administration of the drug is essential. Therefore, there is a need for the development of a new substance having no such side effects and long-term efficacy during long-term administration.

Bone marrow-derived adult stem cells, particularly mesenchymal stem cells (MSCs), are differentiated into various types of cells and tissues, including bone, cartilage, muscle, and fat cells. The differentiation activity of adult stem cells tends to be markedly reduced by disease and aging, and the reduced differentiation activity leads to a decrease in the regenerative activity of bone, cartilage and muscle tissues, accompanied by a weakening of immune function, and infection. Create an environment that can cause various diseases. Therefore, the method of continuously activating the differentiation activity of adult stem cells may be a useful treatment throughout the associated metabolic diseases, bone diseases and diseases associated with aging.

Differentiation of adult stem cells into specific cell types is determined by important transcription factors, which regulate the expression of these genes in response to external signaling and regulate the expression of genes with cell specificity. In particular, differentiation into adipocytes is known to be critically regulated by the peroxisome proliferator-activated receptor γ (PPARγ) transcription factor. The transcription factor promotes or inhibits differentiation into adipocytes by binding to different ligands. It is known that an increase in the activity of the transcription factor promotes differentiation into adipocytes and increases obesity (MacDougald et al. , Annu. Rev. Biochem ., 1995; 64: 345-73; Adams et al., J. Clin. Invest., 1997; 100: 3149-53; Fajas et al., Curr.Opin.Cell.Biol ., 1998; 10: 165-73).

Recently, a transcriptional coactivator with PDZ-binding motif (TAZ) protein that regulates the activity of PPARγ transcription factors has been reported (Kanai et al., Embo. J., 2000; 19: 6778-91). The TAZ protein was cloned into a partner protein that binds to 14-3-3 intracellular protein, and it is known that the 89th serine is bound to 14-3-3 protein by phosphorylation and remains in the cytoplasm (Kanai et al., Embo. J., 2000; 19: 6778-91; Park et al., J. Biol. Chem., 2004; 279: 17384-90). TAZ proteins contain WW domains, coiled-coil domains, and PDZ-binding motifs, suggesting various binding possibilities with other proteins. In particular, the WW domain shows strong binding activity with a peptide array called PPXY, suggesting the possibility of binding to several proteins containing the PPXY motif. In 2003, the binding of RUNX2 (runt-related transcription factor 2) protein, a determinant regulator of osteoblast differentiation, and the WW domain in the TAZ protein was identified, which amplified the target gene expression regulating activity of RUNX2 through osteocalcin. Increasing bone tissue specific gene expression such as (osteocalcin) has been reported to further promote the differentiation of bone tissue (Hong et al., Science 2005; 309: 1074-8). Polyomavirus T antigen is known as a protein that binds to the WW domain of the TAZ protein, but its exact intracellular function is unknown. Furthermore, PPARγ, one of the transcription factors with the PPXY motif, has been identified as a new TAZ binding protein, and this binding has been reported to inhibit adipocyte differentiation activity by PPARγ (Hong et al., Science 2005; 309: 1074-). 8). The mechanism of inhibition of adipocyte differentiation of TAZ protein inhibits the expression of PPARγ target genes having adipocyte specificity by inhibiting DNA binding activity of PPARγ by binding to PPARγ protein and inhibiting transcriptional promoting activity of genes. It is described as. The binding of TAZ protein to RUNX2 and PPARγ is very important for regulating the differentiation of mesenchymal stem cells. In other words, the differentiation of RUNX2 and PPARγ protein and TAZ protein in determining stem cell differentiation promoted osteoblast differentiation by RUNX, while inhibiting adipocyte differentiation by PPARγ (Hong et al. , Science 2005; 309: 1074-8; Deng et al., Front Biosci. 2008; 13: 2001-21; Hong et al., Cell Cycle 2006; 5: 176-9). That is, an important conclusion was drawn that the differentiation fate of mesenchymal stem cells is determined by the expression level of TAZ protein.

In addition, TBX5 (T-box transcription factor 5) is known as a TAZ binding protein, and its binding is understood to play an important role in cardiac and leg development (Murakami et al., Proc. Natl. Acad. Sci. USA. , 2005; 102: 18034-9). It is also suggested to bind and regulate PAX3 protein, which plays an important role in embryonic development (Murakami et al., Biochemical & Biophysical Research Communications 2006; 339: 533-9). In addition, TAZ proteins exhibit activity by binding to various PDZ domain containing proteins via PDZ binding motifs. TTF-1 (thyroid transcription factor-1) is a genetic factor that plays a very important role in the formation of differentiation of lung tissue and regulates the expression of a protein called surfactant protein-C in lung epidermal cells. TAZ protein has been described as a transcriptional promoter that promotes the expression of interfacial protein-C through binding to TTF-1 (Park et al., J. Biol. Chem., 2004; 279: 17384-90). In addition, it has been suggested to regulate the expression of TEF-1 regulatory genes in muscle tissue by binding to transcriptional enhancer factor-1 (TEF-1) proteins (Mahoney et al., Biochem. J., 2005; 388: 217-25).

Apart from the stem cell differentiation regulating function of TAZ protein, the function of TAZ in the migration, invasion and tumorigenesis of MCF7 breast cancer cells is known (Chan et al., Cancer Res., 2008; 68: 2592-8), TAZ protein The incidence of polycystic kidney disease, which forms many cysts in renal cells in the absence of, has been reported in animal models, and approaches to various functions of TAZ proteins have been tried in many ways (Makita et al., Am. J. Physiol. Renal. Physiol. , 2008; 294: F542-53; Tian et al., Molecular & Cellular Biology, 2007; 27: 6383-95). Separately, FGF-2 has been reported as a signal to decrease TAZ protein in osteoblast differentiation (Deng et al., Front Biosci., 2008; 13: 2001-21; Eda et al., Biochemical & Biophysical Research Communications, 2008; 366: 471-5), compared with the importance of the function of the TAZ protein, there is not much research on the regulatory mechanism of the TAZ protein, and further research is required.

In the case of the TAZ protein acting as a transcription factor co-regulatory factor that regulates a transcription factor having DNA binding activity in the cell nucleus, the migration from the cytoplasm to the nucleus is a prerequisite. To date, TAZ protein is known to be able to move to the nucleus by serine dephosphorylation, and inhibition of the binding of TAZ protein to 14-3-3 has been suggested as a way to increase the transfer of TAZ protein to the nucleus. . Therefore, the compound that promotes the migration of the TAZ protein to the nucleus in the cell can be judged to have an effect of inhibiting adipocyte differentiation and promoting osteoblast differentiation by the TAZ protein.

Therefore, the present inventors are studying to find an effective compound for osteoporosis, obesity, diabetes or hyperlipidemia, and the chloroimidazole derivative compound is used for the treatment and prevention of osteoporosis through the regulation of TAZ protein and the treatment of obesity through the inhibition of adipocyte differentiation. It was confirmed that there is an excellent effect and completed the present invention.

An object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of osteoporosis, comprising a chloroimidazole derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of obesity, diabetes or hyperlipidemia, comprising the compound or a pharmaceutically acceptable salt thereof as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating osteoporosis, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

In order to achieve the above another object, the present invention provides a pharmaceutical composition for the prevention or treatment of obesity, diabetes or hyperlipidemia, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

Since the chloroimidazole compound of the present invention or a pharmaceutically acceptable salt thereof modulates the TAZ protein, it can be usefully used for the prevention or treatment of osteoporosis, obesity, diabetes or hyperlipidemia.

The present invention provides a pharmaceutical composition for the prevention or treatment of osteoporosis, obesity, diabetes or hyperlipidemia, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient:

Where

Y is C═O or CH (OH);

X is CH, nitrogen or N-O;

또는

이다.G is hydrogen, CH ₃ , bromine, CH ₂ OH, CH = CH ₂ , CH (OMe) ₂ , CH ₂ N (CH ₂ CH ₃ ) ₂ ,

or

to be.

In particular, in the present invention, the compound of Formula 1 is

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(6-methyl-1-oxy-pyridin-2-yl) -methanone:

;

;

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(6-bromo-1-oxy-pyridin-2-yl) -methanone:

;

;

 {2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(6-bromo-1-oxy-pyridin-2-yl) -methanol:

;

;

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(5-hydroxymethyl-1-oxy-pyridin-2-yl) -methanone:

;

;

6- {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazole-4- represented by the following formula (6): Carbonyl} -1-oxy-pyridine-2-carboxylic acid diethylamide:

;

;

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(6-Dimethoxymethyl-1-oxy-pyridin-2-yl) -methanone:

;

;

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(6-Diethylaminomethyl-1-oxy-pyridin-2-yl) -methanol:

;

;

2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(6-[(diethyloxydoamino) methyl] -1-oxy-pyridin-2-yl) -methanone:

;

;

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(6-methyl-pyridin-2-yl) -methanone:

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(6-bromo-pyridin-2-yl) -methanone:

;

;

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} Pyridin-4-yl-methanone:

;

;

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(5-vinyl-pyridin-2-yl) -methanone:

;

;

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(5-methyl-1-oxy-pyridin-2-yl) -methanone:

;

;

 {2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(1-oxy-pyridin-4-yl) -methanol:

;

;

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} represented by the formula -(4-diethylaminomethyl-phenyl) -methanone:

;

;

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(4-diethylaminomethyl-phenyl) -methanol:

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} represented by the formula -(1-oxy-pyridin-2-yl) -methanol:

;

;

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(5-methyl-pyridin-2-yl) -methanone:

;

;

 {2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} Pyridin-2-yl-methanone:

;

;

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(1-oxy-pyridin-4-yl) -methanone:

;

;

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(1-oxy-pyridin-2-yl) -methanone:

;

;

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(5-methyl-1-oxy-pyridin-2-yl) -methanol:

; 및

; And

{2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -(6-methyl-1-oxy-pyridin-2-yl) -methanol:

It is preferable to select from the group which consists of.

Summarizing the compounds of Chemical Formulas 2 to 24 of the present invention are as follows.

The compounds represented by Formulas 2 to 11 according to the present invention may be prepared and used according to known methods (paper or patent) described in Table 1.

Hereinafter, a method of preparing other compounds will be described in detail.

In one embodiment of preparing the compound of Formula 1, the compound of Formula 1-3 may be prepared via Scheme 1 below.

In Scheme 1, X and G are as defined in Chemical Formula 1.

<Step 1>

화합물(예컨대, 1 내지 3 당량) 및 n-부틸리튬(예컨대, 1 내지 3 당량)을 반응시켜 화학식 1-2의 화합물을 얻을 수 있다. 이때 사용하는 용매로는 테트라히드로퓨란(THF) 또는 디에틸에테르가 바람직하며, 반응조건은 -78℃ 내지 실온에서 3시간 정도 반응시키는 것이 바람직하다.In the coupling reaction according to Step 1 of Scheme 1, an appropriate solvent is used for the compound of Formula 1-1.

A compound of formula 1-2 can be obtained by reacting a compound (eg 1-3 equivalents) and n-butyllithium (eg 1-3 equivalents). At this time, tetrahydrofuran (THF) or diethyl ether is preferable as the solvent to be used, and the reaction conditions are preferably reacted for about 3 hours at -78 ° C to room temperature.

<Step 2>

The active compound of formula 1-2 in the oxidation reaction according to Step 2 of Scheme 1 obtained in step 1 in a suitable solvent, manganese dioxide (MnO ₂₎ (for example, 5 to 10 equivalents) was added and allowed to react for 16 hours formula The compound of -3 can be obtained. At this time, the solvent used is preferably dichloromethane, chloroform, benzene and the like, and the reaction temperature is preferably from room temperature to the boiling point of the solvent.

화합물은 상업적으로 시판되는 것을 사용하거나 제조하여 사용할 수 있다.The compound of Chemical Formula 1-1 used in Scheme 1 may be prepared by a known method (WO 95/21838; US5665738; Bioorg. & Med. Chem., 3 (3), 289, 1995),

The compound can be used commercially available or can be manufactured and used.

In one embodiment of preparing the compound of Formula 1, the compound of Formula 2-2 may be prepared via Scheme 2 below.

 In Scheme 2, Y and G are as defined in Chemical Formula 1.

In the oxidation reaction of Scheme 2, the compound of Formula 2-1 is dissolved in dichloromethane or acetone and m-chloroperoxy benzoic acid ( m- CPBA) (for example, 1 to 3 equivalents) is added, followed by reaction at room temperature for 5 hours. To obtain a compound of formula 2-2 oxidized to pyridyl N-oxide.

In one embodiment of preparing the compound of Formula 1, the compound of Formula 1 may be prepared via Scheme 3 below.

In Scheme 3, Y, X and G are as defined in Chemical Formula 1.

In the deprotection group reaction of Scheme 3, the compound of Formula 3-1 is dissolved in methanol, ethanol or tetrahydrofuran (THF), and then an acid (for example, 1 to 5 equivalents) is added thereto and reacted at room temperature for 10 minutes to 3 hours. The target compound of formula ( 1) from which the protecting group of tetrazole is removed can be obtained. Acid as used here, such as anhydrous hydrochloric acid, hydrochloric acid, p- toluenesulfonic acid, MeSO ₃ H, acetic acid is preferred.

As the pharmaceutically acceptable salt of the compound of Formula 1 of the present invention, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Organic acids and inorganic acids may be used as the free acid, and hydrochloric acid, bromic acid, sulfuric acid, sulfurous acid, phosphoric acid, etc. may be used as the inorganic acid, and citric acid, acetic acid, maleic acid, fumaric acid, gluconic acid, metalsulfonic acid may be used as the organic acid. , Acetic acid, glycolic acid, succinic acid, tartaric acid, 4-toluenesulfonic acid, galacturonic acid, embonic acid, glutamic acid, citric acid, aspartic acid, and the like can be used, and preferably methanesulfonic acid or hydrochloric acid can be used. .

The addition salt according to the present invention is a conventional method, i.e., a compound of formula 1 is dissolved in a water miscible organic solvent such as acetone, methanol, ethanol, or acetonitrile, and an equivalent or excess organic acid is added or an acid aqueous solution of an inorganic acid is added. It may be prepared by addition, followed by precipitation or crystallization, or by evaporation of the solvent or excess acid followed by suction filtration of the dried or precipitated salt.

The present invention includes all of the compounds of Formula 1 and their pharmaceutically acceptable salts as well as the possible solvates, hydrates and stereoisomers that may be prepared therefrom within the scope of the invention.

The compound represented by the formula (1) of the present invention has a use for the prevention or treatment of osteoporosis, obesity, diabetes or hyperlipidemia. Compounds represented by formulas (2) to (24) of the present invention promote the transfer of the transcription factor co-regulatory TAZ protein to the nucleus (Experimental Example 1), and the TAZ protein transferred to the nucleus inhibits the activity of PPARγ through binding to PPARγ. By inhibiting, not only the differentiation of fat cells can be suppressed (Experimental Example 2), but also the differentiation of osteoblasts is promoted by promoting the activity of RUNX2 through binding to the transcription factor RUNX2 (Experimental Example 3). In addition, as a result of experiments with cells, the compounds of the present invention suppress the differentiation into adipocytes, and results in promoting the differentiation of osteoblasts (Experimental Example 4). Therefore, the compounds of the present invention can be usefully used for the prevention or treatment of osteoporosis, obesity, diabetes or hyperlipidemia.

According to the present invention, pharmaceutical compositions comprising a compound of formula 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier can be formulated in a variety of oral or parenteral dosage forms. Formulations for oral administration include, for example, tablets, pills, hard capsules, solutions, suspensions, emulsifiers, syrups, granules, elixirs, etc. Dextrose, sucrose, mannitol, sorbitol, cellulose and / or glycine), lubricants such as silica, talc, stearic acid and its magnesium or calcium salts and / or polyethylene glycols. Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine, optionally starch, agar, alginic acid or its Disintegrants or boiling mixtures such as sodium salts and / or absorbents, colorants, flavors, and sweeteners.

In addition, the pharmaceutical composition comprising the compound represented by the formula (1) as an active ingredient can be administered parenterally, parenteral administration is by a method of injecting subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection. In this case, the compound of Formula 1 or a pharmaceutically acceptable salt thereof is mixed in water with a stabilizer or a buffer to prepare a parenteral formulation, and then prepared as a solution or suspension, which is a unit dosage form of ampoules or vials. It can manufacture.

The compositions may contain sterile and / or preservatives, stabilizers, hydrating or emulsifying accelerators, auxiliaries such as salts and / or buffers for the control of osmotic pressure, and other therapeutically useful substances, and conventional methods of mixing, granulating It may be formulated according to the formulation or coating method.

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples. However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited by the following Examples and Experimental Examples.

In the present invention, the molecular structure of the compounds was confirmed by infrared spectroscopy, nuclear magnetic resonance spectra, mass spectrometry, liquid chromatography, X-ray structure determination, or elemental analysis calculations and actual measurements of representative compounds.

Example 1: {2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -pyridine Preparation of 4-yl-Methanone (Formula 12)

<1-1> 2-butyl-5-chloro-3- {2 '-[1- (1-ethoxy-ethyl) -1H-tetrazol-5-yl] -biphenyl-4-ylmethyl}- Preparation of 3H-imidazole-4-carbaldehyde

2-butyl-5-chloro-3H-imidazole-4-carbaldehyde (3.5 g, 18.7 mmol) was dissolved in 35 mL of N, N-dimethylformamide, followed by potassium carbonate (7.8 g, 56.1 mmol). And 5- (4'-bromomethyl-biphenyl-2-yl) -1- (1-ethoxy-ethyl) -1H-tetrazole (8.0 g, 20.6 mmol) were added and stirred at room temperature for 5 hours. . The reaction solution was diluted with 150 mL of water, extracted with ethyl acetate (100 mL × 2 times), the organic layer was dried over anhydrous sodium sulfate and filtered, and the residue obtained by evaporation and concentration of the solvent under reduced pressure was purified by silica gel column chromatography. Purification by chromatography (n-hexane / ethyl acetate = 2/1) afforded the title compound (8.5 g, yield 92%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.91 ((t, 3H), 1.11 (t, 3H), 1.37 (m, 2H), 1.67 (d, 3H), 1.70 (m, 2H), 2.64 (t , 2H), 3.23 (m, 1H), 3.45 (m, 1H), 5.47 (s, 2H), 5.89 (q, 1H), 6.97 (d, 2H, J = 8.4 Hz), 7.15 (d, 2H, J = 8.4 Hz), 7.40 (dd, 1H, J = 1.6, 7.0 Hz), 7.51 (m, 2H), 7.88 (dd, 1H, J = 1.6, 7.0 Hz), 9.76 (s, 1H, CHO); MS (m / e, M ⁺ ): 492

<1-2> (2-butyl-5- Chloro -3- {2 '-[1- (1- Ethoxy -Ethyl) -1H- Tetrazole -5 days]- Biphenyl -4- Yl methyl } -3H- Imidazole Preparation of 4-yl) -pyridin-4-yl-methanol

4-bromo pyridine (802 mg, 5.07 mmol) was dissolved in 15 mL of tetrahydrofuran (THF) and slowly added n-butyllithium (2.0M solution in cyclohexane) (2.5 mL, 5.07 mmol) at -78 ° C. After dropping, the mixture was stirred for 30 minutes. The compound (1.0 g, 2.03 mmol) obtained in the above <1-1> was diluted with tetrahydrofuran (10 mL) at -78 ° C, and it was slowly added dropwise to the reaction solution. Then, the mixture was slowly raised to room temperature and stirred for 3 hours. The reaction mixture was diluted with 60 mL of water and extracted with ethyl acetate (50 mL × 2 times) and washed with water and brine. The extracted organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by evaporation under reduced pressure. The residue was purified by silica gel column chromatography (n-hexane: ethyl acetate = 2: 1) to give the title compound as a pale yellow solid (750 mg). , Yield 75%) was obtained.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 8.87 (dd, 1H), 8.15 (m, 1H), 7.87 (dd, 1H), 7.50-7.57 (m, 2H), 7.45 (d, 1H), 7.30 ( m, 1H), 7.02-7.05 (m, 2H), 6.70-6.74 (m, 2H), 5.91-5.97 (m, 2H), 5.15-5.23 (m, 2H), 4.06 (s, 1H), 5.73 ( d, 1H), 3.46-3.52 (m, 1H), 3.26-3.32 (m, 1H), 2.52 (t, 2H), 1.62-1.76 (m, 5H), 1.23-1.27 (m, 2H), 1.11 ( t, 3H), 0.84 (t, 3H); MS (m / e, M ⁺ ): 571

<1-3> (2-butyl-5- Chloro -3- {2 '-[1- (1- Ethoxy -Ethyl) -1H- Tetrazole -5 days]- Biphenyl -4-ylmethyl} -3H- Imidazole -4-yl) -pyridin-4-yl- Metathesis  Produce

The compound (1.0 g, 1.75 mmol) obtained in <1-2> was dissolved in 20 mL of dichloromethane, and activated manganese dioxide (1.52 g, 17.51 mmol) was added thereto, followed by stirring for 24 hours. After the reaction was completed, the mixture was filtered through celite, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (n-hexane: ethyl acetate = 2: 1) to give the title compound as a solid ( 796 mg, yield 80%) was obtained.

¹ H-NMR (300 MHz, CDCl ₃ ): d 8.49 (dd, J = 8.4, 2.7 Hz, 1H), 8.24 (d, 1H), 7.87 (dd, J = 8.1, 2.7 Hz, 1H), 7.44-7.58 (m, 3H), 7.41 (d, 2H), 7.18 (d, 2H), 7.02- 7.05 (m, 2H), 5.90 (m, 1H), 5.66 (s, 2H), 3.39-3.50 (m, 1H ), 3.22-3.32 (m, 1H), 2.72 (t, 2H), 1.62-1.76 (m, 5H), 1.31-1.40 (m, 2H), 1.10 (t, 3H), 0.93 (t, 3H); MS (m / e, M ⁺ ): 569

<1-4> {2-butyl-5- Chloro -3- [2 '-(1H- Tetrazole -5 days)- Biphenyl -4- Yl methyl ] -3H- Imidazole -4-yl} -pyridin-4-yl- Metanon  Preparation of Formula 12

The compound (300 mg, 0.52 mmol) obtained in <1-3> was dissolved in 10 mL of methanol-THF (1: 1), and then 3 mL of 3N hydrochloric acid was added thereto, followed by stirring at room temperature for 1 hour. After completion of the reaction, the mixture was neutralized to pH 4-5 with 1 N -sodium hydroxide, and extracted with ethyl acetate (30 mL × 2 times) and washed with water and brine. The extracted organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by evaporation under reduced pressure. The residue was purified by silica gel column chromatography (5% methanol / dichloromethane) to give the title compound as a white solid (238 mg, 91% yield). Obtained.

¹ H-NMR (300 MHz, CDCl ₃ ): d 8.45 (d, J = 5.4, 1H), 8.12 (m, 1H), 7.93 (d, 1H), 7.65 (d, J = 5.4, 1H), 7.50- 7.55 (m, 2H), 7.37 (m, 2H), 7.19 (d, J = 8.1, 2H), 7.00 (d, J = 8.1, 2H), 5.65 (s, 2H), 2.85 (t, 2H), 1.86 (m, 2 H), 1.51 (m, 2 H), 0.96 (t, 3 H); MS (m / e, M ⁺ ): 497

Example  2: {2-butyl-5- Chloro -3- [2 '-(1H- Tetrazole -5 days)- Biphenyl -4- Yl methyl ] -3H- Imidazole -4-yl}-(5-vinyl-pyridin-2-yl)- Metanon  Preparation of Formula 13

<2-1> (2-butyl-5- Chloro -3- {2 '-[1- (1- Ethoxy -Ethyl) -1H- Tetrazole -5 days]- Biphenyl -4- Yl methyl } -3H- Imidazole -4-yl)-(5- Vinylpyridine 2-yl) -Methanol Preparation

In Example <1-2>, except that 2-bromo-5-vinylpyridine (746mg, 4.05 mmol) was used instead of 4-bromo pyridine, and was tested in the same manner as in Example <1-2> The title compound (740 mg, yield 62%) was obtained in the solid state.

¹ H-NMR (300 MHz, CDCl ₃ ) d 8.35 (s, 1H), 7.84 (dd, 1H), 7.44-7.58 (m, 3H), 7.34 (d, 1H), 6.92-6.97 (m, 3H), 6.55-6.66 (m, 3H), 6.01 (s, 1H), 5.91 (q, 1H), 5.73 (d, 1H), 5.42 (s, 1H), 5.31 (dd, 1H), 4.91-5.05 (m, 2H), 3.44-3.50 (m, 1H), 3.23-3.31 (m, 1H), 2.41-2.47 (m, 2H), 1.58-1.72 (m, 5H), 1.24-1.37 (m, 2H), 1.10 ( t, 3H), 0.85 (t, 3H); MS (m / e, M ⁺ ): 597

<2-2> (2-butyl-5- Chloro -3- {2 '-[1- (1- Ethoxy -Ethyl) -1H- Tetrazole -5 days]- Biphenyl -4-ylmethyl} -3H- Imidazole -4-yl)-(5- Vinylpyridine -2 days)- Metathesis  Produce

Example <1-3> is the same as Example <1-3>, except that the compound (730 mg, 1.22 mmol) obtained in <2-1> is used instead of the compound obtained in <1-2>. Experimental method yielded the title compound (580 mg, 80% yield) in the solid state.

¹ H-NMR (300 MHz, CDCl ₃ ) d 8.68 (d, 1H), 7.85-7.90 (m, 2H), 7.72 (d, J = 8.1 Hz, 1H), 7.44-7.55 (m, 2H), 7.39 ( dd, 1H), 7.13 (d, J = 8.4 Hz, 2H), 7.04 (d, J = 8.4 Hz, 2H), 6.78 (dd, 1H), 5.96 (d, 1H), 5.86 (q, 1H), 5.48-5.54 (m, 3H), 3.39-3.47 (m, 1H), 3.19-3.29 (m, 1H), 2.60-2.64 (m, 2H), 1.64-1.74 (m, 5H), 1.33-1.43 (m , 2H), 1.08 (t, 3H), 0.90 (t, 3H); MS (m / e, M ⁺ ): 595

<2-3> {2-butyl-5- Chloro -3- [2'-1H- Tetrazole -5 days)- Biphenyl -4- Yl methyl ] -3H- Imidazole -4-yl)-(5- Vinylpyridine -2 days)- Metanon  Preparation of Formula 13

Example <1-4> except that the compound (150 mg, 0.25 mmol) obtained in Example <2-2> was used instead of the compound obtained in <1-3> in Example <1-4> The experiment was carried out in the same manner as to obtain the title compound (80 mg, 61% yield) in the form of a foamy solid.

¹ H-NMR (300 MHz, CDCl ₃ ) d 8.52 (d, 1H), 7.83-7.90 (m, 2H), 7.68 (d, J = 8.1 Hz, 1H), 7.45-7.57 (m, 2H), 7.36 ( d, J = 7.2 Hz, 1H), 6.98-7.04 (m, 4H), 6.74 (m, 1H), 5.96 (d, 1H), 5.53 (d, 1H), 5.38 (s, 2H), 2.61 (t , 2H), 1.67 (m, 2H), 1.38 (m, 2H), 0.87 (t, 3H); MS (m / e, M ⁺ ): 524

Example  3: {2-butyl-5- Chloro -3- [2 '-(1H- Tetrazole -5 days)- Biphenyl -4- Yl methyl ] -3H- Imidazole -4-yl}-(5-methyl-1- Oxy -Pyridin-2-yl)- Metanon  Preparation of Formula 14

<3-1> (2-butyl-5- Chloro -3- {2 '-[1- (1- Ethoxy -Ethyl) -1H- Tetrazole -5 days]- Biphenyl -4-ylmethyl} -3H- Imidazole -4-yl)-(5- methyl Preparation of -pyridin-2-yl) -methanol

Except for using 2-bromo-5-methyl pyridine (872mg, 5.07 mmol) in place of 4-bromo pyridine in Example <1-2> and was tested in the same manner as in Example <1-2> The title compound (1.0 g, yield 84%) was obtained as a pale yellow solid.

¹ H-NMR (300 MHz, CDCl ₃ ): d 0.86 (t, 3H), 1.11 (t, 3H), 1.31 (m, 2H), 1.62 (m, 2H), 1.72 (d, 3H), 2.21 (s , 3H), 2.41 (t, 2H), 3.26 (m, 1H), 3.46 (m, 1H), 4.96 (d, 2H), 5.49 (s, 1H), 5.91 (m, 1H), 6.00 (s, 1H), 6.64 (m, 2H), 6.91 (d, 1H, J = 7.9 Hz), 6.97 (m, 2H), 7.35 (m, 2H), 7.51 (m, 2H), 7.85 (m, 1H), 8.19 (s, 1 H); MS (m / e, M ⁺ ): 585

<3-2> (2-butyl-5- Chloro -3- {2 '-[1- (1- Ethoxy -Ethyl) -1H- Tetrazole -5 days]- Biphenyl -4-ylmethyl} -3H- Imidazole -4-yl)-(5- methyl -Pyridin-2-yl)- Metathesis  Produce

Example <1-3, except for using the compound (1.0 g, 1.71 mmol) obtained in Example <3-1> instead of the compound obtained in <1-2> in Example <1-3> Experiment was carried out in the same manner as> to give the title compound (975 mg, yield 98%) in the form of a foamy solid.

¹ H-NMR (300 MHz, CDCl ₃ ): d 0.90 (tm 3G), 1.09 (t, 3H), 1.36 (m, 2H), 1.67 (d, 3H), 1.71 (m, 2H), 2.44 (s, 3H), 2.62 (t, 2H), 3.26 (m, 1H), 3.44 (m, 1H), 7.05 (d, 2H, J = 8.0 Hz), 7.14 (d, 2H, J = 8.3 Hz), 7.40 ( dd, 1H, J = 1.6, 7.2 Hz), 7.50 (m, 2H), 7.66 (m, 2H), 7.87 (dd, 1H, J = 1.6, 7.2 Hz), 8.52 (s, 1H); MS (m / e, M ⁺ ): 583

<3-3> (2-butyl-5- Chloro -3- {2 '-[1- (1- Ethoxy -Ethyl) -1H- Tetrazole -5 days]- Biphenyl -4- Yl methyl } -3H- Imidazole -4-yl)-(5- methyl -One- Oxy -Pyridin-2-yl)- Metathesis  Produce

The compound (975 mg, 1.67 mmol) obtained in Example <3-2> was dissolved in 20 mL of dichloromethane, followed by addition of 1.12 g (5.01 mmol, 3eq) of m -chloroperoxybenzoic acid (77%) to 16 Stir for hours. After completion of the reaction mixture was concentrated under reduced pressure and the remaining residue was purified by silica gel column chromatography (methanol: ethyl acetate = 1: 9) to give the title compound (450mg, 45% yield) in the form of a foamy solid.

¹ H-NMR (300 MHz, CDCl ₃ ): d 0.89 (t, 3H), 1.10 (t, 3H), 1.36 (m, 2H), 1.66 (m, 2H), 1.69 (d, 3H), 2.38 (s , 3H), 2.63 (t, 2H), 3.28 (m, 1H), 3.45 (m, 1H), 5.65 (d, 2H), 5.90 (q, 1H), 7.08 (d, 2H, J = 8.2Hz) , 7.18 (d, 4H, J = 9.9 Hz), 7.42 (dd, 1H, J = 1.6, 7.0 Hz), 7.51 (m, 2H), 7.88 (dd, 1H, J = 1.6, 7.0 Hz), 8.09 ( s, 1 H); MS (m / e, M ⁺ ): 599

<3-4> {2-butyl-5- Chloro -3- [2 '-(1H- Tetrazole -5 days)- Biphenyl -4- Yl methyl ] -3H- Imidazole -4-yl}-(5- methyl -One- Oxy -Pyridin-2-yl)- Metanon  Preparation of Formula 14

Example <1-4> except that the compound (50 mg, 0.083 mmol) obtained in Example <3-3> was used instead of the compound obtained in <1-3> in Example <1-4> The experiment was carried out in the same manner as to obtain the title compound (30 mg, yield 70%) in the form of a foamy solid.

¹ H-NMR (300 MHz, CDCl ₃ ): d 0.99 (t, 3H), 1.51 (m, 2H), 1.86 (m, 2H), 2.31 (s, 3H), 2.92 (t, 2H), 5.49 (s , 2H), 7.02 (d, 2H, J = 7.4 Hz), 7.12 (d, 2H, J = 8.1 Hz), 7.30 (m, 2H), 7.41 (d, 1H, J = 6.9 Hz), 7.51 (m , 2H), 7.91 (s, 1H), 8.08 (d, 1H, J = 7.7 Hz); MS (m / e, M ⁺ ): 527

Example  4: {2-butyl-5- Chloro -3- [2 '-(1H- Tetrazole -5 days)- Biphenyl -4- Yl methyl ] -3H- Imidazole Preparation of 4-yl}-(1-oxy-pyridin-4-yl) -methanol (Formula 15)

<4-1> (2-butyl-5- Chloro -3- {2 '-[1- (1- Ethoxy -Ethyl) -1H- Tetrazole -5 days]- Biphenyl -4- Yl methyl } -3H- Imidazole -4-yl)-(1- Oxy Preparation of -pyridin-4-yl) -methanol

Example <3-3, except that the compound (100 mg, 0.175 mmol) obtained in Example <1-2> was used instead of the compound obtained in Example <3-2> in Example <3-3> In the same manner as the> to give the title compound (80 mg, yield 78%) in the form of a foamy solid.

¹ H-NMR (200 MHz, CDCl ₃ ) d 0.86 (t, 3H), 1.10 (t, 3H), 1.25 (m, 4H), 1.70 (d, 3H), 2.42 (m, 2H), 3.30 (m, 1H), 3.35 (m, 1H), 5.02 (m, 2H), 6.00 (d, 1H), 6.65-8.00 (m, 12H); MS (m / e, M ⁺ ): 587

<4-2> {2-butyl-5- Chloro -3- [2 '-(1H- Tetrazole -5 days)- Biphenyl -4- Yl methyl ] -3H- already Zol-4-yl}-(1- Oxy -Pyridin-4-yl) -methanol (Formula 15)

Example <1-4> except that the compound (80 mg, 0.136 mmol) obtained in Example <4-1> was used instead of the compound obtained in <1-3> in Example <1-4> The experiment was carried out in the same manner as to obtain the title compound (63 mg, 90% yield) in the form of a foamy solid.

¹ H-NMR (200 MHz, CDCl ₃ ) d 0.80 (t, 3H), 1.27 (m, 2H), 1.53 (m, 2H), 2.40 (m, 2H), 5.20 (d, 2H), 5.98 (d, 1H), 6.48 (d, 2H), 6.78 (d, 1H), 6.82 (d, 2H), 7.25 (d, 2H), 7.60 (m, 4H), 7.90 (d, 2H); MS (m / e, M ⁺ ): 515

Example  5: {2-butyl-5- Chloro -3- [2 '-(1H- Tetrazole -5 days)- Biphenyl -4- Yl methyl ] -3H- Imidazole -4-yl}-(4-diethylaminomethyl- Phenyl ) - Metanon  Preparation of Formula 16

<5-1> (2-butyl-5- Chloro -3- {2 '-[1- (1- Ethoxy -Ethyl) -1H- Tetrazole -5 days]- Biphenyl -4- Yl methyl } -3H- Imidazole -4-yl)-(4- Diethylaminomethyl - Phenyl )-Preparation of Methanol

Example <1-2>, except that (4-bromo-benzyl) -diethyl-amine (1.23 g, 5.07 mmol) was used instead of 4-bromo pyridine in Example <1-2> The experiment was carried out in the same manner to obtain the title compound (1.0 g, yield 85%) as a foamy solid.

¹ H-NMR (300 MHz, CDCl ₃ ): d 0.85 (t, 3H), 1.02 (m, 6H), 1.10 (t, 3H), 1.30 (m, 2H), 1.61 (m, 2H), 1.70 (d , 3H), 2.41 (t, 2H), 2.50 (q, 4H), 3.25 (m, 1H), 3.46 (m, 1H), 3.50 (d, 2H), 4.95 (d, 2H), 5.91 (q, 1H), 6.10 (s, 1H), 6.71 (d, 2H, J = 7.9 Hz), 7.03 (d, 2H, J = 8.3 Hz), 7.25 (d, 4H, J = 4.7 Hz), 7.40 (m, 1H), 7.51 (m, 2H), 7.90 (m, 1H); MS (m / e, M ⁺ ): 655

<5-2> (2-butyl-5- Chloro -3- {2 '-[1- (1- Ethoxy -Ethyl) -1H- Tetrazole -5 days]- Biphenyl -4-ylmethyl} -3H- Imidazole -4-yl)-(4- Diethylaminomethyl - Phenyl ) - Metathesis  Produce

The compound (500 mg, 0.86 mmol) obtained in <5-1> was dissolved in 15 mL of dichloromethane, and activated manganese dioxide (750 mg, 8.62 mmol) was added thereto, followed by stirring for 24 hours. After the reaction was completed, the mixture was filtered through celite, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (n-hexane: ethyl acetate = 1: 1) to give the title compound as a pale yellow oil (440 mg, Yield 90%) was obtained.

¹ H-NMR (300 MHz, CDCl ₃ ): d 0.92 (t, 3H), 1.04 (t, 6H), 1.08 (t, 3H), 1.40 (m, 2H), 1.65 (d, 3H), 1.75 (m , 2H), 2.52 (q, 4H), 2.66 (t, 2H), 3.21 (m, 1H), 3.43 (m, 1H), 3.62 (s, 2H), 5.47 (s, 2H), 5.83 (q, 1H), 6.98 (d, 2H, J = 8.0 Hz), 7.12 (d, 2H, J = 8.3 Hz), 7.37 (m, 1H), 7.43 (d, 2H, J = 8.0 Hz), 7.50 (m, 2H), 7.69 (d, 2H, J = 8.3 Hz), 7.86 (dd, 1H, J = 1.7, 7.3 Hz); MS (m / e, M ⁺ ): 653

<5-3> {2-butyl-5- Chloro -3- [2 '-(1H- Tetrazole -5 days)- Biphenyl -4- Yl methyl ] -3H- Imidazole -4-yl}-(4- Diethylaminomethyl - Phenyl ) - Metanon  Preparation of Formula 16

Example <1-4> is the same as Example <1-4> except for using the compound (440 mg, 0.76 mmol) obtained in <5-2> instead of the compound obtained in <1-3>. Experiment by the method to give the title compound in the form of a foamy solid (308 mg, 71% yield).

¹ H-NMR (300 MHz, CDCl ₃ ): d 0.95 (t, 3H), 1.33 (t, 6H), 1.45 (m, 2H), 1.80 (m, 2H), 2.80 (t, 2H), 3.06 (q , 4H), 4.07 (s, 2H), 5.30 (s, 2H), 6.72 (d, 2H, J = 8.0 Hz), 7.17 (d, 2H, J = 8.3 Hz), 7.28 (m, 1H), 7.38 (m, 2 H), 7.46 (m, 5 H); MS (m / e, M ⁺ ): 581

Example  6: {2-butyl-5- Chloro -3- [2 '-(1H- Tetrazole -5 days)- Biphenyl -4- Yl methyl ] -3H- Imidazole -4-yl}-(4-diethylaminomethyl- Phenyl ) - Metanon  Preparation of Formula 17

Example <1-4> except that the compound (100 mg, 0.15 mmol) obtained in Example <5-1> was used instead of the compound obtained in <1-3> in Example <1-4> By the same method as in the white solid to obtain the title compound (70mg, 80% yield).

¹ H-NMR (200 MHz, CDCl ₃ ): δ 0.85 (t, 3H), 1.03 (t, 6H), 1.28 (m, 2H), 1.57 (m, 2H), 2.45 (t, 2H), 2.85 ( q, 4H), 4.18 (d, 2H), 5.35 (d, 2H), 6.10 (s, 1H), 6.37 (d, 2H), 6.82 (d, 2H), 7.28 (m, 4H), 7.45 (m , 4H); MS (m / e, M ⁺ ): 583

Example  7: (2-butyl-5- Chloro -3- [2 '-(1H- Tetrazole -5 days)- Biphenyl -4- Yl methyl ] -3H- Imidazole -4-yl}-(1- Oxy -Pyridin-2-yl) -methanol (Formula 18)

<7-1> (2-butyl-5- Chloro -3- {2 '-[1- (1- Ethoxy -Ethyl) -1H- Tetrazole -5 days]- Biphenyl -4- Yl methyl } -3H- Imidazole Preparation of 4-yl) -pyridin-2-yl-methanol

Foamy solids were tested in the same manner as in Example <1-2>, except that 2-bromo pyridine (1.60 g, 10.14 mmol) was used instead of 4-bromo pyridine in Example <1-2>. The title compound (1.15 g, 50% yield) was obtained in the state.

¹ H-NMR (300 MHz, CDCl ₃ ): d 0.85 (t, 3H), 1.12 (t, 3H), 1.34 (m, 2H), 1.65 (m, 2H), 1.73 (d, 3H), 2.43 (t , 2H), 3.28 (m, 1H), 3.48 (m, 1H), 5.03 (m, 2H), 6.00 (m, 1H), 6.04 (s, 1H), 6.63 (m, 2H), 6.94 (m, 2H), 7.07 (m, 2H), 7.36 (d, 1H, J = 7.5 Hz), 7.54 (m, 3H), 7.85 (m, 1H), 8.37 (d, 1H, J = 5.6 Hz); MS (m / e, M ⁺ ): 571

<7-2> (2-butyl-5- Chloro -3- {2 '-[1- (1- Ethoxy -Ethyl) -1H- Tetrazole -5 days]- Biphenyl -4-ylmethyl} -3H- Imidazole -4-yl)-(1- Oxy Preparation of -pyridin-2-yl) -methanol

Example <3- except that the compound (200 mg, 0.35 mmol) obtained in Example <7-1> was used instead of the compound obtained in Example <3-2> in Example <3-3> In the same manner as in the 3>, the title compound (133 mg, yield 65%) was obtained as a white foamy solid.

¹ H-NMR (300 MHz, CDCl ₃ ): d 0.94 (t, 3H), 1.09 (t, 3H), 1.40 (m, 2H), 1.68 (m, 2H), 1.72 (d, 3H), 2.65 (m , 2H), 3.27 (m, 1H), 3.48 (m, 1H), 5.35 (m, 2H), 5.92 (m, 1H), 6.20 (m, 1H), 7.00 (m, 3H), 7.09 (d, 2H, J = 7.7 Hz), 7.21 (m, 2H), 7.37 (m, 1H), 7.52 (m, 2H), 7.88 (M, 1h), 8.17 (m, 1H); MS (m / e, M ⁺ ): 587

<7-3> {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}- Preparation of (1-oxy-pyridin-2-yl) -methanol (Formula 18)

Example <1-4> except that the compound (130 mg, 0.22 mmol) obtained in Example <7-2> was used instead of the compound obtained in <1-3> in Example <1-4> The experiment was carried out in the same manner as to obtain the title compound (95 mg, yield 84%) as a white solid.

¹ H-NMR (200 MHz, CDCl ₃ ): δ 0.96 (t, 3H), 1.46 (m, 2H), 1.85 (m, 2H), 2.80 (t, 2H), 5.35 (m, 2H), 5.96 ( s, 1H), 7.15 (m, 4H), 7.20 (m, 1H), 7.47 (m, 2H), 7.61 (m, 2H), 7.80 (d, 1H, J = 7.6 Hz), 8.03 (m, 2H ); MS (m / e, M ⁺ ): 515

Example 8: 2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6 -[( Diethyloxidoamino ) methyl ]-One- Oxy -Pyridin-2-yl)- Metanon  Preparation of Formula 19

Example <1-4> and Example <1-4> except for using the compound (100 mg, 0.17 mmol) obtained in Example <3-2> instead of the compound obtained in <1-3> The experiment was carried out in the same manner to obtain the title compound (81 mg, yield 92%) as a white solid.

¹ H-NMR (300 MHz, CDCl ₃ ): d 0.95 (t 3H), 1.40 (m, 2H), 1.75 (m, 2H), 2.50 (s, 3H), 2.80 (t, 2H), 5.30 (s, 2H), 6.98 (d, 2H), 7.16 (d, 2H), 7.25 (m, 1H), 7.35 (m, 2H), 7.45 (m, 4H); MS (m / e, M ⁺ ): 511

Example 9: {2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -pyridine Preparation of 2-yl-Methanone (Formula 20)

<9-1> (2-butyl-5-chloro-3- {2 '-[1- (1-ethoxy-ethyl) -1H-tetrazol-5-yl] -biphenyl-4-ylmethyl} Preparation of -3H-imidazol-4-yl) -pyridin-2-yl-methanone

Example <1-3> except that the compound (1.14 g, 1.99 mmol) obtained in Example <7-1> was used instead of the compound obtained in <1-2> in Example <1-3> The experiment was carried out in the same manner as to obtain the title compound (1.07 g, yield 95%) in the form of a foamy solid.

¹ H-NMR (300 MHz, CDCl ₃ ): d 0.91 (t, 3H), 1.10 (t, 3H), 1.40 (m, 2H), 1.68 (d, 3H), 1.74 (m, 2H), 2.64 (t , 2H), 3.25 (m, 1H), 3.45 (m, 1H), 5.51 (s, 2H), 5.89 (q, 1H), 7.06 (d, 2H, J = 8.0 Hz), 7.16 (d, 2H, J = 8.2 Hz), 7.48 (m, 4H), 7.73 (d, 1H, J = 7.0 Hz), 7.87 (m, 2H), 8.71 (d, 1H, J = 4.0 Hz); MS (m / e, M ⁺ ): 569

<9-2> {2-butyl-5- Chloro -3- [2 '-(1H- Tetrazole -5 days)- Biphenyl -4- Yl methyl ] -3H- Imidazole -4-yl} -pyridin-2-yl- Metanon  Preparation of Formula 19

Example <1-4> is the same as Example <1-4>, except that the compound (300 mg, 0.52 mmol) obtained in <9-1> is used instead of the compound obtained in <1-3>. The experiment was carried out to obtain the title compound (220 mg, yield 86%) as a white solid.

¹ H-NMR (300 MHz, CDCl ₃ ): d 0.92 (t, 3H), 1.47 (m, 2H), 1.75 (m, 2H), 2.71 (t, 2H), 5.48 (s, 2H), 7.05 (d , 2H, J = 8.1Hz), 7.18 (d, 2H, J = 8.1Hz), 7.48 (m, 4H), 7.75 (d, 1H, J = 7.1Hz), 7.86 (m, 2H), 8.73 (d , 1H, J = 4.2 Hz); MS (m / e, M ⁺ ): 497

Example 10 {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-( Preparation of 1-oxy-pyridin-4-yl) -methanone (Formula 21)

<10-1> (2-butyl-5-chloro-3- {2 '-[1- (1-ethoxy-ethyl) -1H-tetrazol-5-yl] -biphenyl-4-ylmethyl} Preparation of -3H-imidazol-4-yl)-(1-oxy-pyridin-4-yl) -methanone

Example <3-3, except for using the compound (150 mg, 0.26 mmol) obtained in Example <1-3> instead of the compound obtained in Example <3-2> in Example <3-3> The experiment was carried out in the same manner as> to obtain the title compound (88 mg, yield 58%) as a white foamy solid.

¹ H-NMR (200 MHz, CDCl ₃ ): d 0.92 (t, 3H), 1.08 (t, 3H), 1.49 (m, 4H), 1.68 (d, 3H), 2.71 (t, 2H), 3.25 (m , 1H), 3.46 (m, 1H), 5.41 (s, 2H), 5.88 (q, 1H), 6.92 (d, 2H), 7.11 (d, 2H), 7.32 (m, 1H), 7.48 (m, 2H), 7.55 (d, 2H), 7.85 (m, 1H), 8.20 (d, 2H); MS (m / e, M ⁺ ): 585

<10-2> {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} Preparation of-(1-oxy-pyridin-4-yl) -methanone (Formula 21)

Example <1-4> except that the compound (85 mg, 0.15 mmol) obtained in Example <10-1> was used instead of the compound obtained in <1-3> in Example <1-4> By the same method as in the white solid to obtain the title compound (63mg, 82% yield).

¹ H-NMR (300 MHz, CDCl ₃ ): d 0.92 (t, 3H), 1.85 (m, 2H), 1.51 (m, 2H), 2.73 (t, 2H), 5.41 (s, 2H), 6.93 (d , 2H), 7.10 (d, 2H), 7.36 (m, 1H), 7.48 (m, 2H), 7.57 (d, 2H), 7.85 (m, 1H), 8.23 (d, 2H); MS (m / e, M ⁺ ): 513

Example  11: (2-butyl-5- Chloro -3- [2 '-(1H- Tetrazole -5 days)- Biphenyl -4- Yl methyl ] -3H- Imidazole -4-yl}-(1- Oxy -Pyridin-2-yl)- Metanon  Preparation of Formula 22

<11-1> (2-butyl-5-chloro-3- {2 '-[1- (1-ethoxy-ethyl) -1H-tetrazol-5-yl] -biphenyl-4-ylmethyl} Preparation of -3H-imidazol-4-yl)-(1-oxy-pyridin-2-yl) -methanone

Example <3 except for using the compound (1.10 g, 1.93 mmol) obtained in Example <9-1> instead of the compound obtained in Example <3-2> in Example <3-3> The experiment was carried out in the same manner as in -3>, to obtain the title compound (688 mg, 61% yield) as a white foamy solid.

¹ H-NMR (300 MHz, CDCl ₃ ): d 0.89 (t, 3H), 1.10 (t, 3H), 1.37 (m, 2H), 1.66 (m, 2H), 1.69 (d, 3H), 2.64 (t , 2H), 3.27 (m, 1H), 3.46 (m, 1H), 5.66 (m, 2H), 5.90 (m, 1H), 7.09 (d, 2H, J = 8.2 Hz), 7.18 (d, 2H, J = 8.2 Hz), 7.30 (m, 1H), 7.39 (m, 3H), 7.51 (m, 2H), 7.88 (dd, 1H, J = 1.5, 7.5 Hz), 8.24 (m, 1H); MS (m / e, M ⁺ ): 585

<11-2> {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}- Preparation of (1-oxy-pyridin-2-yl) -methanone (Formula 22)

Example <1-4> except that the compound (210 mg, 0.36 mmol) obtained in Example <11-1> was used instead of the compound obtained in <1-3> in Example <1-4> By the same method as in the white solid to obtain the title compound (172mg, 93% yield).

¹ H-NMR (300 MHz, CDCl ₃ ): d 1.01 (t, 3H), 1.43 (m, 2H), 2.97 (t, 2H), 7.09 (d, 2H, J = 8.3 Hz), 7.17 (d, 2H , J = 8.3 Hz), 7.43 (d, 3H, J = 7.2 Hz), 7.55 (m, 3H), 8.10 (d, 1H, J = 6.1 Hz), 8.15 (d, 1H, J = 7.2 Hz); MS (m / e, M ⁺ ): 513

Example  12: {2-butyl-5- Chloro -3- [2 '-(1H- Tetrazole -5 days)- Biphenyl -4- Yl methyl ] -3H- Imidazole -4-yl}-(5-methyl-1- Oxy Preparation of -pyridin-2-yl) -methanol (Formula 23)

<12-1> (2-butyl-5- Chloro -3- {2 '-[1- (1- Ethoxy -Ethyl) -1H- Tetrazole -5 days]- Biphenyl - 4- Yl methyl } -3H- Imidazole -4-yl)-(5- methyl -One- Oxy Preparation of -pyridin-2-yl) -methanol

Example <3- except that the compound (150 mg, 0.25 mmol) obtained in Example <3-1> was used instead of the compound obtained in Example <3-2> in Example <3-3> By the same method as 3> to give the title compound (90 mg, yield 60%) as a white foamy solid.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 8.02 (s, 1H), 7.88 (m, 1H), 7.51 (m, 2H), 7.38 (d, J = 7.2 Hz, 1H), 7.08 (d, J = 8.0 Hz, 2H), 6.99 (d, J = 8.0 Hz, 2H), 6.84 (m, 1H), 6.40 (m, 1H), 6.13 (s, 1H), 5.91 (m, 1H), 5.31 (s, 2H), 3.47 (m, 1H), 3.27 (m, 1H), 2.61 (t, 2H), 2.25 (s, 3H), 1.77 (d, 3H), 1.38 (m, 2H), 1.10 (t, 3H) ), 0.90 (t, 3 H); MS (m / e, M ⁺ ): 601

<12-2> {2-butyl-5- Chloro -3- [2 '-(1H- Tetrazole -5 days)- Biphenyl -4- Yl methyl ] -3H- Imidazole -4-yl}-(5- methyl -One- Oxy Preparation of -pyridin-2-yl) -methanol (Formula 23)

Example <1-4> except for using the compound (80 mg, 0.133 mmol) obtained in Example <12-1> instead of the compound obtained in <1-3> in Example <1-4> The experiment was carried out in the same manner as to obtain the title compound (63 mg, yield 90%) as a white solid.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 7.96 (d, J = 7.5 Hz, 1H), 7.80 (s, 1H), 7.59 (m, 2H), 7.48 (m, 2H), 7.21 (d, J = 8.3 Hz, 1H), 7.12 (d, J = 8.0 Hz, 2H), 7.06 (d, J = 8.0 Hz, 2H), 5.94 (s, 1H), 5.41 (d, J = 16.3 Hz, 1H), 5.22 (d, J = 16.3 Hz, 1H), 2.72 (t, 2H), 2.22 (s, 3H), 1.73 (m, 2H), 1.41 (m, 2H), 0.93 (t, 3H); MS (m / e, M ⁺ ): 529

Example  13: {2-butyl-5- Chloro -3- [2 '-(1H- Tetrazole -5 days)- Biphenyl -4- Yl methyl ] -3H- Imidazole -4-yl}-(6-methyl-1- Oxy Preparation of -pyridin-2-yl) -methanol (Formula 24)

<13-1> (2-butyl-5- Chloro -3- {2 '-[1- (1- Ethoxy -Ethyl) -1H- Tetrazole -5 days]- Biphenyl -4- Yl methyl } -3H- Imidazole -4-yl)-(6- methyl Preparation of -pyridin-2-yl) -methanol

Experiment in the same manner as in Example <1-2>, except that 2-bromo-6-methyl pyridine (0.53 g, 3.08 mmol) was used instead of 4-bromo pyridine in Example <1-2> To give the title compound as a foamy solid (327 mg, yield 55%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 7.85 (d, J = 7.7 Hz, 1H), 7.53 (m, 2H), 7.37 (m, 2H), 6.93 (m, 3H), 6.82 (d, J = 7.4 Hz, 1H), 6.61 (t, 2H), 5.98 (s, 1H), 5.89 (t, 1H), 5.75 (s, 1H), 4.96 (m, 2H), 3.46 (m, 1H), 2.43 ( s, 3H), 2.38 (t, 2H), 1.71 (m, 3H), 1.61 (m, 2H), 1.28 (m, 2H), 1.11 (t, 3H), 0.85 (t, 3H); MS (m / e, M ⁺ ): 585

<13-2> (2-butyl-5- Chloro -3- {2 '-[1- (1- Ethoxy -Ethyl) -1H- Tetrazole -5 days]- Biphenyl -4- Yl methyl } -3H- Imidazole -4-yl)-(6- methyl -One- Oxy Preparation of -pyridin-2-yl) -methanol

Example <3-3> except that the compound (300 mg, 0.51 mmol) obtained in <13-1> was used instead of the compound obtained in Example <3-2> in Example <3-3> The experiment was carried out in the same manner as to obtain the title compound (163 mg, yield 53%) as a white foamy solid.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 7.88 (m, 1H), 7.56 (m, 2H), 7.38 (m, 2H), 7.15 (m, 2H), 7.09 (m, 1H), 6.99 (d, J = 8.0 Hz, 2H), 6.84 (m, 1H), 6.40 (m, 1H), 6.19 (s, 1H), 5.88 (m, 1H), 5.32 (s, 2H), 3.43 (m, 1H), 3.22 (m, 1H), 2.54 (t, 2H), 2.52 (s, 3H), 1.67 (d, 3H), 1.35 (m, 2H), 1.08 (t, 3H), 0.88 (t, 3H); MS (m / e, M ⁺ ): 601

<13-3> {2-butyl-5- Chloro -3- [2 '-(1H- Tetrazole -5 days)- Biphenyl -4- Yl methyl ] -3H- Imidazole -4-yl}-(6- methyl -One- Oxy Preparation of -pyridin-2-yl) -methanol (Formula 24)

Example <1-4> is the same as Example <1-4>, except that the compound (150 mg, 0.25 mmol) obtained in <13-2> is used instead of the compound obtained in <1-3>. The experiment was carried out to obtain the title compound (112 mg, yield 85%) as a white solid.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 7.87 (d, J = 7.7 Hz, 1H), 7.58 (d, J = 7.1 Hz, 1H), 7.49 (m, 2H), 7.45 (d, J = 7.1 Hz , 1H), 7.26 (m, 1H), 7.21 (m, 1H), 7.09 (m, 4H), 5.93 (s, 1H), 5.39 (d, J = 16.6 Hz, 1H), 5.27 (d, J = 16.6 Hz, 1H), 2.65 (t, 2H), 2.28 (s, 3H), 1.69 (m, 2H), 1.38 (m, 2H), 0.89 (t, 3H); MS (m / e, M ⁺ ): 529

Substituents of the compounds of the present invention are shown in Table 2 below.

Compound (Formula) Y X G 2 C = O (2) -N-O (3) -Me 3 C = O (2) -N-O (3) -Br 4 CH-OH (2) -N-O (3) -Br 5 C = O (2) -N-O (4) -CH ₂ OH 6 C = O (2) -N-O (3) -CON (Et) ₂ 7 C = O (2) -N-O (3) -CH (OMe) ₂ 8 CH-OH (2) -N-O (3) -CH ₂ N (Et) ₂ 9 C = O (2) -N-O (3) -CH ₂ NO (Et) ₂ 10 C = O (2) -N (3) -Me 11 C = O (2) -N (3) -Br 12 C = O (4) -N H 13 C = O (2) -N (4) -CH = CH ₂ 14 C = O (2) -N-O (4) -Me 15 CH-OH (4) -N-O H 16 C = O CH (4) -CH ₂ N (Et) ₂ 17 CH-OH CH (4) -CH ₂ N (Et) ₂ 18 CH-OH (2) -N-O H 19 C = O (2) -N (4) -Me 20 C = O (2) -N H 21 C = O (4) -N-O H 22 C = O (2) -N-O H 23 CH-OH (2) -N-O (4) -Me 24 CH-OH (2) -N-O (3) -Me

Various pharmacological effects were evaluated by performing the following experiments on the compound of Formula 1 according to the present invention.

Experimental Example 1: Verification of the effect of the compound of the present invention on the migration of the TAZ protein to the nucleus

To investigate the effect of the compound of formula 1 according to the present invention on the migration of intracellular TAZ protein to the nucleus, a vector (pEGFP-TAZ) and a cell in which a green fluorescence protein (GFP) is linked to the TAZ protein Nuclear specific histone protein (RFP-H2B) expression vector to which a red fluorescence protein (RFP) is linked to identify the position of the inner nucleus was prepared. Specifically, the pEGFP-TAZ vector obtained a full-length TAZ cDNA by the PCR method, and then cloned into the pEGFP vector (Invitrogen, Carlsbad, CA, USA), the RFP-H2B expression vector RFP expression of full-length histone H2B cDNA Inserted into a vector (Clontech Laboratories, Inc., Palo Alto, Calif., USA), cloned and used.

Cos7 cells (ATCC, Manassas, VA) (5x10 ³ cells / well) were placed and stabilized in 96-well plates, and then pEGFP-TAZ and RFP-H2B vectors were introduced into cells by effectene (Qiagen) method. 48 hours after the introduction of the vector, the cells were treated with the compounds of the formulas 2 to 24 according to the present invention at a concentration of 10 μM each. 30 minutes after the treatment, the BD pathway high-content bioimager (BD bioscience) was used to check the movement of the green fluorescent nucleus in the cells in real time. Quantification programs (BD IPLab ™ for Pathway and BD ™ Image Data Explorer) were used to quantify the expression of GFP signals in the nucleus of cells expressing RFP signals.

The degree of GFP signal expression in the nucleus of the TAZ protein expressed in the control cells not treated with the compound was set to 100%, and the change by the compound of the present invention was expressed in%.

The measurement results are shown in Table 3 below.

Effect of Compounds of the Invention on Transfer of TAZ Proteins to the Nucleus compound
Formula 2 3 4 5 6 7 8 9 10 11 Promotion effect
(%) 150 149 149 156 158 153 141 150 125 126 compound
Formula 12 13 14 15 16 17 18 20 21 23 Promotion effect
(%) 155 156 130 140 165 146 151 122 113 151 compound
Formula 24 Promotion effect
(%) 129

As a result, 16 of the 23 compounds of the present invention were found to significantly increase the expression in the nucleus of the TAZ protein by more than 130%. Thus, the compounds of the present invention were expected to affect adipocyte differentiation and osteoblast differentiation by promoting the migration of TAZ proteins into the nucleus.

Experimental Example 2: Verification of the effects of the compounds of the present invention on TAZ activity that inhibits PPARγ function

To investigate the effects of compounds of the present invention on TAZ activity that inhibits PPARγ function, 293T cells (ATCC) were plated in 48-well plates (1 × 10 ⁵ cells / well) and stabilized, followed by PPARγ, TAZ expression vectors. AP2-luc (adipose fatty acid binding protein 2 promoter linked to luciferase) and a pCMVβ vector were introduced (see Hong et al., Science 2005; 309: 1074-8). In order to observe the effect on the transcriptional activity of the target gene according to the expression of PPARγ or TAZ protein, aP2-luc reporter gene was introduced into the cell together, and all of the pCMVβ vectors were introduced into the same cell and β- Galactosidase activity was measured and corrected for luciferase activity values.

After 24 hours, the cells were treated with compounds of formulas 2 to 24 according to the present invention at a concentration of 10 μM each and further incubated for 24 hours. Cell proteins were extracted using NP-40 containing lysate from the cultured cells, and luciferase activity was measured (Promega, Sunnyvale, CA, USA). The increase in aP2 promoter activity by PPARγ was reduced by TAZ expression, the reduced value in cells not treated with the compound according to the present invention was set to 100%, and the effect of activity inhibition by the compound according to the present invention was expressed in%. It was. The measurement results are shown in Table 4 below.

Influence of Compounds of the Invention on TAZ Activity Inhibiting PPARγ Function compound
Formula 2 3 4 5 6 7 8 9 12 13 Inhibitory Effect (%) 51.3 37.1 55.2 55.4 10.6 42.0 54.6 22.2 40.2 63.9 compound
Formula 14 15 16 17 18 19 22 Inhibitory Effect (%) 34.7 39.8 57.7 42.0 48.7 50.5 30.1

As shown in Table 4, 17 of the 23 compounds of the present invention showed an additional inhibitory effect, in particular, in the case of compounds of Formulas 2, 4, 5, 8, 13, 16 and 19, 50% or more It showed a strong inhibitory effect. Therefore, the compounds of the present invention inhibit the adipocyte differentiation activity of PPARγ through the binding between TAZ and PPARγ, it can be seen that it is effective in preventing or treating obesity.

Experimental Example 3: Verification of the effects of the compounds of the present invention on TAZ activity promoting RUNX2 function

To investigate the effects of the compounds of the present invention on TAZ activity promoting RUNX2 function, 6 × OSE-luc (osteocalcin-specific element linked to luciferase, six copies of RUNX2-binding) with RUNX2 and TAZ expression vectors in 293T cells site in the osteocalcin promoter linked to luciferase) (see Hong et al., Science 2005; 309: 1074-8). The expression of RUNX2 protein increased the binding of OSE promoter to luciferase activity.

After 24 hours, the cells were treated with compounds of Formulas 2 to 24 at 10 μM concentrations and incubated for 24 hours. After extracting the cellular protein in the same manner as in Experimental Example 2, the reporter gene analysis was performed to quantify the effect of the compounds on the RUNX2 promoting function of TAZ. In this case, in order to compare the transformation efficiency, pCMVβ vector expressing β-galactosidase was introduced into 293T cells, and then β-galactosidase activity was measured and corrected. An increase in osteocalcin promoter activity by RUNX2 was observed, and the level in the cells not treated with the compound was set to 100%. It was confirmed that the average increase of 570% by the additional expression of the TAZ protein, it was determined that the compound having a higher activity than that, it has an additional activity promoting effect. The measurement results are shown in Table 5 below.

Effect of compounds on TAZ activity (570%) on promoting RUNX2 function compound
Formula 2 3 4 5 6 7 8 9 12 13 Promotion effect (%) 681.6 521.2 597.8 477.8 579.8 873.6 575.7 1066.5 1215.7 451.1 compound
Formula 14 15 16 17 18 19 23 24 Promotion effect (%) 528.7 659.7 626.2 696.5 990.5 170 424.4 410

As shown in Table 5, 11 of 23 compounds showed further promoting activity, and in particular, the compounds of Formulas 9, 12, and 18 showed further promoting activity. Therefore, the compounds of the present invention promote the differentiation of RUNX2 into osteoblasts through the binding between TAZ and the transcription factor RUNX2, it can be seen that it is effective in the prevention or treatment of osteoporosis.

Experimental Example 4: Analysis of the effect of the compounds of the present invention on adipocyte or osteoblast differentiation

In order to observe the effect of the compounds of the present invention on the differentiation of adipocytes or osteoblasts, 3T3-L1 cells were induced to differentiate into adipocytes, and C3H10T1 / 2 cells were used to induce differentiation into osteoblasts.

Induction of Differentiation of 3T3-L1 Cells into Adipocytes

In order to induce differentiation of 3T3-L1 cells into adipocytes, 3T3-L1 adipocytes (ATCC CL-173) were suspended in DMEM medium containing 10% Calf serum, followed by 24-well plates ( Incubate for 48 hours to grow confluent at the bottom of 3 × 10 ⁴ cells / well). Thereafter, 2 μM rosiglitazone, 5 μg / mL insulin and 1 μM dexamethasone were added to the medium to which 10% fetal bovine serum (FBS) was added to induce differentiation into adipocytes. After 48 hours, the medium was replaced with DMEM medium containing 5 μg / mL insulin and 10% fetal bovine serum, again 48 hours later, with DMEM medium containing 10% fetal bovine serum and freshly changed medium every 48 hours. Adipose cell differentiation was observed. Compounds according to the invention were further fed when the medium was changed. After 8 days of induction of differentiation, the cells were fixed with 10% formalin and then oil-red O staining was performed to check the fat produced in the cells.

As a result, as shown in Fig. 1, the differentiation into adipocytes was reduced by the 16 compounds tested, and in particular, the differentiation into adipocytes was significantly reduced by the compounds of Formulas 2, 4, 6 and 12. Could confirm.

<4-2> Induction of Differentiation of C3H10T1 / 2 Cells into Osteoblasts

To induce differentiation of C3H10T1 / 2 cells into osteoblasts, C3H10T1 / 2 cells (ATCC CCL 226) were diluted in α-MEM medium containing 10% fetal calf serum and 96-well plates (1 × 10 ⁴ cells). / Well) incubated for 48 hours, and then differentiation into osteoblasts was observed while changing medium at 48 hour intervals. Compounds according to the invention were treated at a concentration of 10 μM on the start of differentiation. Twenty days after the induction of differentiation, bone intracellular production was confirmed by Alizarin red staining in response to increased calcium. Differentiated cells were fixed with 70% ethanol and then stained with Alizarin Red S solution. The better the differentiation into osteoblasts, the stronger the red light, so you can compare the degree of differentiation through the degree of staining.

As a result, as shown in Figure 2, it was found that the differentiation into osteoblasts was promoted by the fifteen compounds tested, and in particular, the formulas 2, 3, 5, 6, 8, 12, 13, 16 and Compound 18 has been shown to significantly promote differentiation into osteoblasts.

Therefore, the compounds according to the present invention can inhibit the differentiation of adipocytes, promote the differentiation of osteoblasts, can be effectively used to inhibit obesity or to improve osteoporosis.

Figure 1 shows the effect of the compounds of the present invention on adipocyte differentiation (compounds B-1 to B-23 each represent a compound of formula 2 to formula 24).

Figure 2 shows the effect of the compounds of the present invention on osteoblast differentiation (compounds B-1 to B-23 each represent a compound of formula 2 to formula 24).

Claims (4)

A pharmaceutical composition for preventing or treating osteoporosis, comprising the compound represented by the following Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient: <Formula 1>

Where Y is C═O or CH (OH); X is CH, nitrogen or N-O; G is hydrogen, CH ₃ , bromine, CH ₂ OH, CH = CH ₂ , CH (OMe) ₂ , CH ₂ N (CH ₂ CH ₃ ) ₂ ,

or

to be. The compound of claim 1 wherein said compound is (1) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6 -Methyl-1-oxy-pyridin-2-yl) -methanone; (2) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6 -Bromo-1-oxy-pyridin-2-yl) -methanone; (3) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6 -Bromo-1-oxy-pyridin-2-yl) -methanol; (4) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(5 -Hydroxymethyl-1-oxy-pyridin-2-yl) -methanone; (5) 6- {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazole-4-carbonyl} -1-oxy-pyridine-2-carboxylic acid diethylamide; (6) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6 -Dimethoxymethyl-1-oxy-pyridin-2-yl) -methanone; (7) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6 -Diethylaminomethyl-1-oxy-pyridin-2-yl) -methanol; (8) 2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6- [(Diethyloxydoamino) methyl] -1-oxy-pyridin-2-yl) -methanone; (9) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6 -Methyl-pyridin-2-yl) -methanone; (10) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6 -Bromo-pyridin-2-yl) -methanone; (11) {2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -pyridine -4-yl-methanone; (12) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(5 -Vinyl-pyridin-2-yl) -methanone; (13) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(5 -Methyl-1-oxy-pyridin-2-yl) -methanone; (14) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(1 -Oxy-pyridin-4-yl) -methanol; (15) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(4 -Diethylaminomethyl-phenyl) -methanone; (16) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(4 -Diethylaminomethyl-phenyl) -methanol; (17) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(1 -Oxy-pyridin-2-yl) -methanol; (18) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(5 -Methyl-pyridin-2-yl) -methanone; (19) {2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -pyridine- 2-yl-methanone; (20) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(1 -Oxy-pyridin-4-yl) -methanone; (21) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-( 1-oxy-pyridin-2-yl) -methanone; (22) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(5 -Methyl-1-oxy-pyridin-2-yl) -methanol; And (23) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6 -Methyl-1-oxy-pyridin-2-yl) -methanol Composition selected from the group consisting of. A pharmaceutical composition for preventing or treating obesity, diabetes or hyperlipidemia, comprising the compound represented by the following Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient: <Formula 1>

Where Y is C═O or CH (OH); X is CH, nitrogen or N-O; G is hydrogen, CH ₃ , bromine, CH ₂ OH, CH = CH ₂ , CH (OMe) ₂ , CH ₂ N (CH ₂ CH ₃ ) ₂ ,

or

to be. The compound of claim 3 wherein said compound is (1) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6 -Methyl-1-oxy-pyridin-2-yl) -methanone; (2) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6 -Bromo-1-oxy-pyridin-2-yl) -methanone; (3) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6 -Bromo-1-oxy-pyridin-2-yl) -methanol; (4) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(5 -Hydroxymethyl-1-oxy-pyridin-2-yl) -methanone; (5) 6- {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazole-4-carbonyl} -1-oxy-pyridine-2-carboxylic acid diethylamide; (6) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6 -Dimethoxymethyl-1-oxy-pyridin-2-yl) -methanone; (7) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6 -Diethylaminomethyl-1-oxy-pyridin-2-yl) -methanol; (8) 2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6- [(Diethyloxydoamino) methyl] -1-oxy-pyridin-2-yl) -methanone; (9) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6 -Methyl-pyridin-2-yl) -methanone; (10) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6 -Bromo-pyridin-2-yl) -methanone; (11) {2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -pyridine- 4-yl-methanone; (12) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(5 -Vinyl-pyridin-2-yl) -methanone; (13) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(5 -Methyl-1-oxy-pyridin-2-yl) -methanone; (14) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(1 -Oxy-pyridin-4-yl) -methanol; (15) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(4 -Diethylaminomethyl-phenyl) -methanone; (16) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(4 -Diethylaminomethyl-phenyl) -methanol; (17) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(1 -Oxy-pyridin-2-yl) -methanol; (18) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-( 5-methyl-pyridin-2-yl) -methanone; (19) {2-Butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl} -pyridine- 2-yl-methanone; (20) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(1 -Oxy-pyridin-4-yl) -methanone; (21) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(1 -Oxy-pyridin-2-yl) -methanone; (22) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(5 -Methyl-1-oxy-pyridin-2-yl) -methanol; And (23) {2-butyl-5-chloro-3- [2 '-(1H-tetrazol-5-yl) -biphenyl-4-ylmethyl] -3H-imidazol-4-yl}-(6 -Methyl-1-oxy-pyridin-2-yl) -methanol Composition selected from the group consisting of.

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