NOVEL COMPOUND AND PHARMACEUTICAL COMPOSITION COMPRISING SAME AS ACTIVE INGREDIENT

Abstract

The present invention relates to a novel compound and a pharmaceutical composition comprising the same as an active ingredient. The novel compound, which is obtained by means of carrying out chemical modification in a mother structure showing excellent AMP-activated protein kinase (AMPK) activation efficacy and in silico binding capacity, enables effective prevention or treatment of degenerative neurological disorders by means of effectively inducing AMP-activated protein kinase enzyme activation.

Claims

1. A compound represented by the following Formula 1: ##STR00005## in the Formula 1, L.sub.1 to L.sub.2 are each independently selected from the group consisting of C.sub.3 to C.sub.40 cycloalkylene, C.sub.6 to C.sub.60 arylene, heteroarylene having 5 to 60 nuclear atoms; X and Y are each independently selected from the group consisting of deuterium, halogen, cyano, nitro, sulfonyl, C.sub.1 to C.sub.10 alkylsulfonyl, azide, hydroxy, C.sub.1 to C.sub.40 alkyl, C.sub.2 to C.sub.40 alkenyl, C.sub.1 to C.sub.40 alkoxy, unsubstituted or substituted C.sub.6 to C.sub.60 aryloxy, unsubstituted or substituted C.sub.3 to C.sub.40 cycloalkyl, unsubstituted or substituted heterocycloalkyl having 3 to 20 nuclear atoms, unsubstituted or substituted C.sub.6 to C.sub.60 aryl, unsubstituted or substituted heteroaryl having 5 to 60 nuclear atoms, and NRR; R and R are each independently selected from the group consisting of hydrogen, C.sub.1 to C.sub.10 alkyl, C.sub.6 to C.sub.60 aryl, C.sub.3 to C.sub.40 cycloalkyl, C.sub.6 to C.sub.60 arylsulfonyl, heteroaryl having 5 to 60 nuclear atoms; and n is selected from integers of 0 to 5.

2. The compound according to claim 1, wherein L.sub.1 and L.sub.2 are C.sub.6 to C.sub.60 arylene.

3. The compound according to claim 1, wherein L.sub.1 and L.sub.2 are phenylene; n is an integer of 1 or 2; and X and Y are the same as or different from each other and are each independently selected from the group consisting of sulfonyl, C.sub.1 to C.sub.10 alkylsulfonyl, C.sub.1 to C.sub.40 alkoxy, NRR, hydroxy, C.sub.6 to C.sub.60 aryloxy, and unsubstituted or substituted heterocycloalkyl having 3 to 20 nuclear atoms.

4. The compound according to claim 3, wherein X is NRR; and R and R are the same as or different from each other and are each independently hydrogen or a substituent of the following Formula 2: ##STR00006## in the formula, R.sub.1 is selected from the group consisting of hydrogen, deuterium, halogen, and nitro.

5. The compound according to claim 3, wherein X is the following Formula 3: ##STR00007## in the formula, R.sub.2 is hydrogen or hydroxy.

6. The compound according to claim 3, wherein Y is C.sub.1 to C.sub.6 alkoxy.

7. The compound according to claim 3, wherein Y is the following Formula 4: ##STR00008##

8. The compound according to claim 1, wherein the compound is selected from the group consisting of the following compounds: ##STR00009## ##STR00010## ##STR00011##

9. A method for preparing the compound according to claim 1, comprising: 1) a step of mixing an acetophenone derivative and a benzaldehyde derivative with an organic solvent, and performing stirring; and 2) a step of extracting the reactant of the step 1) using an organic solvent wherein the acetophenone derivative is 3-aminoacetophenone or 4-aminoacetophenone.

9. (canceled)

10. The method according to claim 9, wherein the benzaldehyde derivative is selected from any one of the compound 2,4-dimethoxybenzaldehyde, 2,5-dimethoxybenzaldehyde, 4-methoxybenzaldehyde, 2-methoxy-4-((tetrahydro-2H-pyran-2-yl)oxy)benzaldehyde, or 4-(piperidin-1-yl)benzaldehyde.

11. The method according to claim 9, further comprising, after the step 2), a step of adding a substituted alkylsulfonyl chloride-based compound and a base, and performing mixing and stirring.

12. A method for preventing or treating degenerative neurological diseases, comprising administering to a subject in need thereof, the compound of claim 1 as an active ingredient.

13. The method according to claim 12, wherein the degenerative neurological disease is one or more selected from the group consisting of stroke, palsy, dementia, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), Pick's disease and Creutzfeldt-Jakob disease.

14. The method according to claim 12, wherein the degenerative neurological disease is a disease caused by brain neuron damage resulting from amyloid-beta peptide-induced stress.

15. The method according to claim 12, wherein the composition induces the AMP-activated protein kinase (AMPK) enzyme activation.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0046] FIG. 1 illustrates an in silico schematic diagram according to an embodiment of the present invention.

[0047] FIG. 2 illustrates results obtained by performing virtual selection for a preparation example according to an embodiment of the present invention.

[0048] FIG. 3 illustrates results obtained by performing a kinase assay according to an embodiment of the present invention.

[0049] FIG. 4 illustrates a circular pool for Morris water maze test according to an embodiment of the present invention.

[0050] FIG. 5 illustrates results obtained by performing Morris water maze test according to an embodiment of the present invention.

[0051] FIG. 6 illustrates results obtained by performing Morris water maze test according to an embodiment of the present invention.

[0052] FIG. 7 illustrates results obtained by performing Morris water maze test according to an embodiment of the present invention.

[0053] FIG. 8 illustrates results obtained by performing a probe test according to an embodiment of the present invention.

[0054] FIG. 9 illustrates results obtained by performing a probe test according to an embodiment of the present invention.

[0055] FIG. 10 illustrates results obtained by performing a passive avoidance test according to an embodiment of the present invention.

[0056] FIG. 11 illustrates results obtained by performing a rotarod test according to an embodiment of the present invention.

[0057] FIG. 12 illustrates results obtained by performing a vertical pole test according to an embodiment of the present invention.

[0058] FIG. 13 illustrates results obtained by measuring activity of acetylcholinesterase (AchE) according to an embodiment of the present invention.

[0059] FIG. 14 illustrates results obtained by measuring acetylcholine (Ach) according to an embodiment of the present invention.

[0060] FIG. 15 illustrates results obtained by performing TUNEL assay according to an embodiment of the present invention.

[0061] FIG. 16 illustrates results obtained by performing TUNEL assay according to an embodiment of the present invention.

[0062] FIG. 17 illustrates analysis of degree of membrane permeability for Preparation Example 6.

[0063] FIG. 18 illustrates results obtained by performing half-life analysis for Preparation Example 6.

DETAILED DESCRIPTION OF INVENTION

[0064] The present invention provides a novel compound, (E)-4-chloro-N-(4-(3-(2,5-dimethoxyphenyl)acryloyl)phenyl)benzenesulfonamide, and provides a pharmaceutical composition for preventing or treating degenerative neurological diseases, comprising the compound as an active ingredient.

[0065] Hereinafter, the present invention will be described in more detail by way of examples. These examples are only for describing the present invention in more detail, and it will be apparent to those skilled in the art that according to the gist of the present invention, the scope of the present invention is not limited by these examples.

[Experimental Example 1] Selection of Drug Candidates with AMP-Activated Protein Kinase Activation Efficacy for Treatment of Degenerative Neurological Diseases

[0066] In order to select candidates for the treatment of degenerative neurological diseases, drug candidates capable of binding to AMP-activated protein kinase were selected.

[0067] Selection of the candidates was made by identifying, through in silico and in vitro screening methods, candidates capable of binding to a regulatory site on the AMP-activated protein kinase enzyme, from a chemical library.

[0068] As illustrated in FIG. 1, CNa87, which is the form that can best bind to the AMP-activated protein kinase enzyme, was finally selected.

[0069] Using CNa87 as a mother body, Preparation Examples 1 to 15, which are candidates obtained by modifying CNa87 with a group that is bulkier than a hydroxyl group so as to have further enhanced binding ability to the AMP-activated protein kinase enzyme, were synthesized by the following methods.

[Preparation Examples 1 to 15] Synthesis of Candidates

[0070] Preparation Examples 1 to 15 in Table 1 below were prepared by the following Method 1 or Method 2.

TABLE-US-00001 TABLE 1 Preparation example Compound name Preparation (E)-1-(4-Aminophenyl)-3- Example 1 (2,4-dimethoxyphenyl)prop-2-en-1-one Preparation (E)-1-(4-Aminophenyl)-3- Example 2 (2,5-dimethoxyphenyl)prop-2-en-1-one Preparation (E)-1-(3-Aminophenyl)-3- Example 3 (4-methoxyphenyl)prop-2-en-1-one Preparation (E)-1-(3-Aminophenyl)-3-(4- Example 4 hydroxy-2-methoxyphenyl)prop-2-en-1-one Preparation (E)-1-(3-Aminophenyl)-3- Example 5 (2,5-dimethoxyphenyl)prop-2-en-1-one Preparation (E)-4-Chloro-N-(4-(3-(2,5- Example 6 dimethoxyphenyl)acryloyl)phenyl)benzenesulfonamide Preparation (E)-4-Chloro-N-(3-(3-(4- Example 7 methoxyphenyl)acryloyl)phenyl)benzenesulfonamide Preparation (E)-4-Chloro-N-(3-(3-(2,5- Example 8 dimethoxyphenyl)acryloyl)phenyl)benzenesulfonamide Preparation (E)-4-Chloro-N-((4-chlorophenyl)sulfonyl)-N-(3-(3-2,5- Example 9 dimethoxyphenyl)acryloyl)phenyl)benzenesulfonamide Preparation (E)-1-(4-Aminophenyl)-3-(4- Example 10 (piperidin-1-yl)phenyl)prop-2-en-1-one Preparation (E)-1-(3-Aminophenyl)-3-(4- Example 11 (piperidin-1-yl)phenyl)prop-2-en-1-one Preparation (E)-1-(4-Hydroxyphenyl)-3-(4- Example 12 (piperidin-1-yl)phenyl)prop-2-en-1-one Preparation (E)-1-(2,4-Dihydroxyphenyl)-3-(4- Example 13 (piperidin-1-yl)phenyl)prop-2-en-1-one Preparation (E)-3-(4-Hydroxy-2-methoxyphenyl)-1- Example 14 (4-(piperazin-1-yl)phenyl)prop-2-en-1-one Preparation (E)-3-(4-Hydroxyphenyl)-1-(4- Example 15 (4-methylpiperazin-1-yl)phenyl)prop-2-en-1-one

[Method 1]

[0071] An acetophenone derivative (1 equivalent), a benzaldehyde derivative (1 equivalent), and NaOH (1 equivalent) were added to an ethanol solvent and stirring was performed at room temperature. Water was added to the reaction mixture and extraction with ethyl acetate was performed. The organic solvent layer was collected and washed once again with water. Anhydrous MgSO.sub.4 was added thereto and dehydration was performed. Then, the solvent was distilled off under reduced pressure, and the remaining residue was purified by silica gel chromatography, to prepare the compound.

[Method 2]

[0072] An acetophenone derivative (1 equivalent), 4-((tetrahydro-2H-pyran-2-yl)oxy))benzaldehyde derivative (1 equivalent), and NaOH (1 equivalent) were added to an ethanol solvent and stirring was performed at room temperature. 4M HCl was added to the reaction mixture and stirred for 20 more minutes. Then, water was added thereto and extraction with ethyl acetate was performed. The organic solvent layer was collected and washed once again with water. Anhydrous MgSO.sub.4 was added thereto and dehydration was performed. Then, the solvent was distilled off under reduced pressure, and the remaining residue was purified by silica gel chromatography, to prepare the compound.

[Preparation Example 1] (E)-1-(4-Aminophenyl)-3-(2,4-dimethoxyphenyl)prop-2-en-1-one)

[0073] According to the above Method 1, 4-aminoacetophenone (0.30 g, 2.22 mmol), 2,4-dimethoxybenzaldehyde (0.37 g, 2.22 mmol), and NaOH (0.09 g, 2.22 mmol) were used and purification by silica gel chromatography (developing solvent: ethyl acetate/n-hexane=1:2 1:1) was performed, to obtain yellow Preparation Example 1 (0.15 g, 23.0%). R.sub.f 0.33 (ethyl acetate/n-hexane=1:1); .sup.1H-NMR (400 MHz, CDCl.sub.3) 3.85 (s, 3H), 3.89 (s, 3H), 6.47 (d, J=2.0 Hz, 1H), 6.52 (dd, J=8.4, 2.4 Hz, 1H), 6.69 (d, J=8.4 Hz, 2H), 7.55 (d, J=15.6 Hz, 1H), 7.56 (d, J=8.8 Hz, 1H), 7.92 (d, J=8.4 Hz, 2H), 8.02 (d, J=15.6 Hz, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 55.6, 55.7, 98.7, 105.5, 114.1, 117.8, 120.7, 129.4, 130.8, 131.1, 139.0, 150.9, 160.4, 162.8, 189.1 ppm.

[Preparation Example 2] (E)-1-(4-Aminophenyl)-3-(2,5-dimethoxyphenyl)prop-2-en-1-one)

[0074] According to the above Method 1, 4-aminoacetophenone (0.50 g, 3.70 mmol), 2,5-dimethoxybenzaldehyde (0.62 g, 3.70 mmol), and NaOH (0.15 g, 3.70 mmol) were purified by silica gel chromatography (developing solvent: ethyl acetate/n-hexane=1:2 1:1), to obtain yellow Preparation Example 2 (0.66 g, 62.5%). R.sub.f 0.36 (ethyl acetate/n-hexane=1:1); .sup.1H-NMR (400 MHz, CDCl.sub.3) 3.81 (s, 3H), 3.85 (s, 3H), 4.19 (br s, 2H), 6.69 (d, J=8.8 Hz, 2H), 6.86 (d, J=8.8 Hz, 1H), 6.91 (dd, J=8.8, 2.8 Hz, 1H), 7.16 (d, J=2.8 Hz, 1H), 7.58 (d, J=15.6 Hz, 1H), 7.92 (d, J=8.8 Hz, 2H), 8.04 (d, J=15.6 Hz, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 56.0, 56.3, 112.7, 113.9, 114.1, 116.8, 123.3, 125.2, 128.9, 131.3, 138.6, 151.2, 153.4, 153.7, 188.8 ppm.

[Preparation Example 3] (E)-1-(3-Aminophenyl)-3-(4-methoxyphenyl)prop-2-en-1-one

[0075] According to the above Method 1, 3-aminoacetophenone (1.00 g, 7.40 mmol), 4-methoxybenzaldehyde (1.00 g, 7.40 mmol), and NaOH (0.30 g, 7.40 mmol) were used and purification by silica gel chromatography (developing solvent: ethyl acetate/n-hexane=1:2 1:1) was performed, to obtain orange Preparation Example 3 (0.83 g, 62.5%). R.sub.f 0.40 (ethyl acetate/n-hexane=1:1); .sup.1H-NMR (400 MHz, CDCl.sub.3) 3.83 (br s, 2H), 3.85 (s, 3H), 6.88 (ddd, J=8.0, 2.4, 0.8 Hz, 1H), 6.93 (d, J=8.8 Hz, 2H), 7.27 (dd, J=8.0, 7.6 Hz, 1H), 7.31 (dd, J=2.0, 1.6 Hz, 1H), 7.36 (d, J=15.6 Hz, 1H), 7.38 (ddd, J=7.6, 1.6, 0.8 Hz, 1H), 7.59 (d, J=8.8 Hz, 2H), 7.76 (d, J=15.6 Hz, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 56.0, 56.3, 112.7, 113.9, 114.1, 116.8, 123.3, 125.2, 128.9, 131.3, 138.6, 151.2, 153.4, 153.7, 188.8 ppm.

[Preparation Example 4] (E)-1-(3-Aminophenyl)-3-(4-hydroxy-2-methoxyphenyl)prop-2-en-1-one)

[0076] According to the above Method 2, 3-aminoacetophenone (0.40 g, 2.96 mmol), 2-methoxy-4-((tetrahydro-2H-pyran-2-yl)oxy)benzaldehyde (0.70 g, 2.96 mmol), and NaOH (0.12 g, 2.96 mmol) were used and purification by silica gel chromatography (developing solvent: ethyl acetate/n-hexane=1:1) was performed, to obtain orange Preparation Example 4 (0.28 g, 35.1%). R.sub.f 0.25 (ethyl acetate/n-hexane=1:1); .sup.1H-NMR (400 MHz, CDCl.sub.3) 3.71 (s, 3H), 6.30 (d, J=2.4 Hz, 1H), 6.34 (dd, J=8.4, 2.0 Hz, 1H), 6.72 (ddd, J=8.0, 2.4, 0.8 Hz, 1H), 7.08 (dd, J=8.4, 8.0 Hz, 1H), 7.13 (d, J=2.4 Hz, 1H), 7.16 (ddd, J=7.6, 7.6, 0.8 Hz, 1H), 7.29 (d, J=15.6 Hz, 1H), 7.32 (d, J=8.4 Hz, 1H), 7.85 (d, J=15.6 Hz, 1H), 9.35 (br s, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 55.3, 99.1, 108.2, 114.1, 115.3, 118.1, 118.7, 119.3, 129.1, 130.6, 139.8, 140.3, 147.0, 60.5, 161.4, 191.1 ppm.

[Preparation Example 5] (E)-1-(3-Aminophenyl)-3-(2,5-dimethoxyphenyl)prop-2-en-1-one)

[0077] According to the above Method 1, 4-aminoacetophenone (0.50 g, 3.70 mmol), 2,5-dimethoxybenzaldehyde (0.62 g, 3.70 mmol), and NaOH (0.15 g, 3.70 mmol) were used and purification by silica gel chromatography (developing solvent: ethyl acetate/n-hexane=1:3) was performed, to obtain yellow Preparation Example 5 (0.27 g, 25.4%). R.sub.f 0.62 (ethyl acetate/n-hexane=1:1); .sup.1H-NMR (400 MHz, CDCl.sub.3) 3.81 (s, 3H), 3.86 (s, 3H), 6.87 (d, J=8.8 Hz, 1H), 6.88 (ddd, J=8.4, 2.0, 0.8 Hz, 1H), 6.94 (dd, J=8.8, 2.8 Hz, 1H), 7.16 (d, J=2.8 Hz, 1H), 7.27 (dd, J=8.0, 8.0 Hz, 1H), 7.31 (dd, J=2.8, 2.8 Hz, 1H), 7.38 (ddd, J=8.0, 2.0, 1.6 Hz, 1H), 7.53 (d, J=15.6 Hz, 1H), 8.06 (d, J=15.6 Hz, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 56.1, 56.3, 112.7, 113.9, 114.7, 117.4, 119.1, 119.4, 123.6, 124.8, 129.6, 139.8, 140.0, 147.0, 153.5, 153.7, 191.4 ppm.

[Preparation Example 6] (E)-4-Chloro-N-(4-(3-(2,5-dimethoxyphenyl)acryloyl)phenyl)benzenesulfonamide

[0078] 4-Chlorobenzenesulfonyl chloride (0.75 g, 3.54 mmol) was added to a CH.sub.2Cl.sub.2 solution in which the Preparation Example 2 (0.67 g, 2.36 mmol) and triethylamine (TEA, 0.26 g, 2.60 mmol) are dissolved, and stirring was performed at room temperature for 24 hours. Water was added to the reaction mixture, and extraction with ethyl acetate was performed. The organic solvent layer was collected and washed with saturated NaHCO.sub.3. Anhydrous MgSO.sub.4 was added thereto and dehydration was performed. Then, the solvent was distilled off under reduced pressure, and the remaining residue was purified by silica gel chromatography (developing solvent: ethyl acetate/n-hexane=1:2), to obtain yellow Preparation Example 6 (0.55 g, 50.9%). R.sub.f 0.18 (ethyl acetate/n-hexane=1:1); .sup.1H-NMR (400 MHz, DMSO-d6) 3.74 (s, 3H), 3.80 (s, 3H), 6.84 (d, J=8.8 Hz, 1H), 6.88 (dd, J=8.8, 2.4 Hz, 1H), 7.16 (d, J=2.4 Hz, 1H), 7.20 (d, J=8.4 Hz, 2H), 7.40 (d, J=8.4 Hz, 2H), 7.54 (d, J=15.6 Hz, 1H), 7.74 (d, J=8.4 Hz, 2H), 7.85 (d, J=8.4 Hz, 2H), 7.91 (d, J=15.6 Hz, 1H), 10.58 (s, 1H); .sup.13C-NMR (100 MHz, DMSO-d6) 55.2, 55.6, 112.1, 112.7, 117.0, 118.0, 121.6, 123.5, 128.0, 128.1, 128.7, 129.4, 132.9, 137.8, 138.3, 141.5, 152.5, 152.9, 187.9 ppm.

[Preparation Example 7] (E)-4-Chloro-N-(3-(3-(4-methoxyphenyl)acryloyl)phenyl)benzenesulfonamide

[0079] N-(3-Acetylphenyl)-4-chlorobenzenesulfonamide (0.10 g, 0.32 mmol), 4-methoxybenzaldehyde (0.04 g, 0.32 mmol), and NaOH (0.03 g, 0.80 mmol) was added to an ethanol solvent and stirring was performed at room temperature for 72 hours. A dilute hydrochloric acid aqueous solution was added to the reaction mixture, and extraction with ethyl acetate was performed. The organic solvent layer was collected and washed with water. Anhydrous MgSO.sub.4 was added thereto and dehydration was performed. Then, the solvent was distilled off under reduced pressure. Subsequently, the remaining residue was purified by silica gel chromatography (developing solvent: ethyl acetate/n-hexane=1:3 2:1), to obtain yellow Preparation Example 7 (0.01 g, 6.5%). .sup.1H-NMR (400 MHz, CDCl.sub.3) 3.84 (s, 3H), 6.92 (d, J=8.8 Hz, 2H), 7.28 (d, J=15.6 Hz, 1H), 7.33 (dd, J=8.0, 7.6 Hz, 1H), 7.36 (d, J=8.8 Hz, 2H), 7.41 (ddd, J=8.0, 2.0, 1.2 Hz, 1H), 7.57 (d, J=8.8 Hz, 2H), 7.68 (ddd, J=7.6, 1.6, 1.2 Hz, 1H), 7.71-7.78 (m, 4H), 9.25 (s, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 55.6, 114.6, 119.5, 121.1, 124.9, 125.0, 127.6, 128.8, 129.4, 129.7, 130.5, 137.

[Preparation Example 8] (E)-4-Chloro-N-(3-(3-(2,5-dimethoxyphenyl)acryloyl)phenyl)benzenesulfonamide

[0080] 4-Chlorobenzenesulfonyl chloride (0.09 g, 0.43 mmol) was added to a CH.sub.2Cl.sub.2 solution in which the Preparation Example 5 (0.12 g, 0.43 mmol) and triethylamine (0.03 g, 2.60 mmol) are dissolved, and stirring was performed at room temperature for 24 hours. Water was added to the reaction mixture, and extraction with ethyl acetate was performed. The organic solvent layer was collected and washed with saturated NaHCO.sub.3. Anhydrous MgSO.sub.4 was added thereto and dehydration was performed. Then, the solvent was distilled off under reduced pressure, and the remaining residue was purified by silica gel chromatography (developing solvent: ethyl acetate/n-hexane=1:3), to obtain yellow Preparation Example 8 (0.08 g, 37.3%). R.sub.f 0.33 (ethyl acetate/n-hexane=1:1); .sup.1H-NMR (400 MHz, CDCl.sub.3) 3.82 (s, 3H), 3.88 (s, 3H), 6.88 (d, J=8.8 Hz, 1H), 6.96 (dd, J=8.4, 2.8 Hz, 1H), 7.15 (d, J=2.8 Hz, 1H), 7.40 (d, J=8.8 Hz, 2H), 7.41-7.43 (m, 2H), 7.55 (d, J=16.0 Hz, 1H), 7.73 (d, J=8.8 Hz, 2H), 7.71-7.72 (m, 1H), 7.76-7.79 (m, 1H), 8.08 (d, J=16.0 Hz, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3)56.1, 56.3, 112.7, 114.5, 117.9, 121.6, 122.8, 124.4, 125.5, 125.8, 128.9, 129.7, 130.0, 137.1, 137.7, 139.9, 140.0, 141.6, 153.7, 153.8, 190.3 ppm.

[Preparation Example 9] (E)-4-Chloro-N-((4-chlorophenyl)sulfonyl)-N-(3-(3-(2,5-dimethoxyphenyl)acryloyl)phenyl)benzenesulfonamide

[0081] 4-Chlorobenzenesulfonyl chloride (0.25 g, 1.17 mmol) was added to a CH.sub.2C.sub.12 solution in which the Preparation Example 5 (0.22 g, 0.78 mmol) and triethylamine (0.22 g, 2.12 mmol) are dissolved, and stirring was performed at room temperature for 24 hours. Water was added to the reaction mixture, and extraction with ethyl acetate was performed. The organic solvent layer was collected and washed with saturated NaHCO.sub.3. Anhydrous MgSO.sub.4 was added thereto and dehydration was performed. Then, the solvent was distilled off under reduced pressure, and the remaining residue was purified by silica gel chromatography (developing solvent: ethyl acetate/n-hexane=1:4), to obtain yellow Preparation Example 9 (0.20 g, 39.8%). R.sub.f 0.77 (ethyl acetate/n-hexane=1:1); .sup.1H-NMR (400 MHz, CDCl.sub.3) 3.83 (s, 3H), 3.86 (s, 3H), 6.89 (d, J=9.2 Hz, 1H), 6.97 (dd, J=9.2, 3.2 Hz, 1H), 7.13 (d, J=2.8 Hz, 1H), 7.19 (dd, J=8.4, 2.0 Hz, 1H), 7.42 (d, J=16.0 Hz, 1H), 7.52 (d, J=8.4 Hz, 1H), 7.54 (d, J=8.8 Hz, 4H), 7.66 (dd, J=1.6, 1.6 Hz, 1H), 7.89 (d, J=8.8 Hz, 4H), 8.05 (d, J=16.0 Hz, 1H), 8.09 (d, J=8.0 Hz, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 56.1, 56.3, 112.7, 114.3, 117.9, 122.9, 124.3, 129.8, 129.9, 130.3, 130.6, 131.6, 134.6, 135.2, 137.8, 140.2, 141.4, 141.8, 153.7, 153.8, 189.9 ppm.

[Preparation Example 10] (E)-1-(4-Aminophenyl)-3-(4-(piperidin-1-yl)phenyl)prop-2-en-1-one)

[0082] According to the above Method 1, 4-aminoacetophenone (0.40 g, 2.96 mmol), 4-(piperidin-1-yl)benzaldehyde (0.56 g, 2.96 mmol), and NaOH (0.12 g, 2.96 mmol) were used and purification by silica gel chromatography (developing solvent: MeOH:CHCl3=1:19) was performed, to obtain yellow Preparation Example 10 (0.28 g, 30.9%). R.sub.f 00.43 (ethyl acetate/n-hexane=1:1); .sup.1H-NMR (400 MHz, CDCl.sub.3) 1.62-1.71 (m, 6H), 3.29 (t, J=5.2 Hz, 4H), 4.10 (br s, 2H), 6.69 (d, J=8.8 Hz, 2H), 6.89 (d, J=8.8 Hz, 2H), 7.37 (d, J=15.6 Hz, 1H), 7.53 (d, J=8.8 Hz, 2H), 7.75 (d, J=15.6 Hz, 1H), 7.95 (d, J=8.8 Hz, 2H).

[Preparation Example 11] (E)-1-(3-Aminophenyl)-3-(4-(piperidin-1-yl)phenyl)prop-2-en-1-one)

[0083] According to the above Method 1,3-aminoacetophenone (0.40 g, 2.96 mmol), 4-(piperidin-1-yl)benzaldehyde (0.56 g, 2.96 mmol), and NaOH (0.12 g, 2.96 mmol) were used and purification by silica gel chromatography (developing solvent: MeOH:CHCl.sub.3=1:19) was performed, to obtain yellow Preparation Example 11 (0.45 g, 49.6%). R.sub.f 0.47 (ethyl acetate/n-hexane=1:1); .sup.1H-NMR (400 MHz, CDCl.sub.3) 1.61-1.72 (m, 6H), 3.31 (t, J=5.6 Hz, 4H), 3.80 (br s, 2H), 6.86 (ddd, J=8.0, 2.4, 0.8 Hz, 1H), 6.89 (d, J=8.8 Hz, 2H), 7.28 (dd, J=8.0, 8.0 Hz, 1H), 7.30 (d, J=15.6 Hz, 1H), 7.31 (dd, J=2.0, 2.0 Hz, 1H), 7.37 (ddd, J=8.8, 1.2, 1.2 Hz, 1H), 7.53 (d, J=8.8 Hz, 2H), 7.75 (d, J=15.6 Hz, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 24.6, 25.7, 49.3, 114.7, 115.0, 118.4, 119.0, 119.1, 124.7, 129.5, 130.4, 140.3, 145.4, 146.9, 153.4, 191.1 ppm.

[Preparation Example 12] (E)-1-(4-Hydroxyphenyl)-3-(4-(piperidin-1-yl) henyl) prop-2-en-1-one)

[0084] According to the above Method 2, 1-(4-((tetrahydro-2H-pyran-2-yl)oxy)phenyl)ethan-1-one (0.50 g, 2.27 mmol), 4-(piperidin-1-yl)benzaldehyde (0.43 g, 2.27 mmol), and NaOH (0.09 g, 2.27 mmol) were used and purification by silica gel chromatography (developing solvent: ethyl acetate/n-hexane=1:3 1:1) was performed, to obtain orange Preparation Example 12 (0.24 g, 33.7%). R.sub.f 0.17 (ethyl acetate/n-hexane=1:3); .sup.1H-NMR (400 MHz, CDCl.sub.3) 1.58-1.65 (m, 6H), 3.25 (t, J=5.6 Hz, 4H), 6.84 (d, J=8.8 Hz, 2H), 6.87 (d, J=8.8 Hz, 2H), 7.32 (d, J=15.6 Hz, 1H), 7.48 (d, J=8.8 Hz, 2H), 7.68 (d, J=15.6 Hz, 1H), 7.90 (d, J=8.8 Hz, 2H), 9.34 (s, 1H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 24.4, 25.5, 49.2, 114.9, 115.6, 117.8, 124.8, 130.1, 130.6, 130.9, 144.2, 153.1, 161.7, 189.0 ppm.

[Preparation Example 13] (E)-1-(2,4-Dihydroxyphenyl)-3-(4-(piperidin-1-yl)phenyl)prop-2-en-1-one)

[0085] According to the above Method 2, 1-(4-((tetrahydro-2H-pyran-2-yl)oxy)phenyl)ethan-1-one (0.50 g, 2.12 mmol), 4-(piperidin-1-yl)benzaldehyde (0.40 g, 2.12 mmol), and Ba(OH).sub.28H.sub.2O (0.73 g, 2.33 mmol) were used and purification by silica gel chromatography (developing solvent: ethyl acetate/n-hexane=1:1) was performed, to obtain orange Preparation Example 13 (0.12 g, 16.8%). R.sub.f 0.66 (ethyl acetate/n-hexane=1:1); .sup.1H-NMR (400 MHz, CDCl.sub.3) 1.58-1.65 (m, 6H), 3.27 (t, J=5.6 Hz, 4H), 6.37-6.40 (m, 2H), 6.84 (d, J=8.8 Hz, 2H), 7.34 (d, J=15.2 Hz, 1H), 7.49 (d, J=8.8 Hz, 2H), 7.73 (d, J=8.4 Hz, 1H), 7.76 (d, J=15.2 Hz, 1H), 9.64 (s, 1H), 13.6 (s, 1H).

[Preparation Example 14] (E)-3-(4-Hydroxy-2-methoxyphenyl)-1-(4-(piperazin-1-yl)phenyl)prop-2-en-1-one)

[0086] According to the above Method 2,1-(4-((tetrahydro-2H-pyran-2-yl)oxy)phenyl)ethan-1-one (0.50 g, 2.45 mmol), 2-methoxy-4-((tetrahydro-2H-pyran-2-yl)oxy)benzaldehyde (0.58 g, 2.45 mmol), and NaOH (0.20 g, 4.90 mmol) were used, and the solid obtained after removal of the solvent was treated with a mixed solvent of ethyl acetate/n-hexane. The resulting solid was filtered and dried in vacuo, to obtain orange Preparation Example 14 (0.28 g, 33.8%). .sup.1H-NMR (400 MHz, DMSO-d6) 2.89 (dd, J=7.2, 3.2 Hz, 4H), 3.26 (dd, J=7.2, 4.0 Hz, 4H), 3.82 (s, 3H), 6.40 (dd, J=8.8, 2.0 Hz, 1H), 6.41 (d, J=1.6 Hz, 1H), 6.89 (d, J=8.8 Hz, 2H), 7.51 (d, J=15.6 Hz, 1H), 7.55 (d, J=8.8 Hz, 1H), 7.87 (d, J=15.6 Hz, 1H), 7.90 (d, J=8.8 Hz, 2H); .sup.13C-NMR (100 MHz, DMSO-d6) 45.1, 47.5, 55.1, 98.7, 107.9, 112.9, 114.7, 118.0, 127.7, 129.7, 129.8, 137.7, 153.7, 159.7, 161.2, 186.8 ppm.

[Preparation Example 15] (E)-3-(4-Hydroxyphenyl)-1-(4-(4-methylpiperazin-1-yl)phenyl)prop-2-en-1-one

[0087] According to the above Method 2,1-(4-((tetrahydro-2H-pyran-2-yl)oxy)phenyl)ethan-1-one (0.50 g, 2.29 mmol), 4-((tetrahydro-2H-pyran-2-yl)oxy)benzaldehyde (0.47 g, 2.29 mmol), and NaOH (0.09 g, 2.29 mmol) were used, and the solid obtained after removal of the solvent was treated with a mixed solvent of ethyl acetate/n-hexane. The resulting solid was filtered and dried in vacuo, to obtain orange Preparation Example 15 (0.70 g, 94.8%). .sup.1H-NMR (400 MHz, CDCl.sub.3) 2.33 (s, 3H), 2.55 (t, J=5.2 Hz, 4H), 3.37 (t, J=5.2 Hz, 4H), 6.86 (d, J=8.8 Hz, 2H), 6.89 (d, J=8.8 Hz, 2H), 7.38 (d, J=15.6 Hz, 1H), 7.49 (d, J=8.8 Hz, 2H), 7.72 (d, J=15.6 Hz, 1H), 7.95 (d, J=8.8 Hz, 2H); .sup.13C-NMR (100 MHz, CDCl.sub.3) 46.2, 47.4, 54.8, 113.7, 116.2, 118.8, 126.8, 128.7, 130.3, 130.6, 143.7, 154.0, 159.7, 188.4 ppm.

[Experimental Example 2] In Silico 3D Docking Test

[0088] Binding ability of the Preparation Examples 1 to 15 to the AMP-activated protein kinase (AMPK) enzyme was identified.

[0089] Using, as a control, the CNa87 molecule which is a mother body of the candidates, 3D docking experiments were carried out on the Preparation Examples 1 to 15, and the results are shown in Table 2 and FIG. 2.

[0090] As shown in Table 2 and FIG. 2, as compared with the control having a docking score of 5.5818, the Preparation Example 6 obtained by substitution with 4-chlorobenzenesulfonamide group exhibited the highest docking score of 8.2507; and the Preparation Examples 7 and 8 also exhibited a docking score which is about 1 or higher than the control.

TABLE-US-00002 TABLE 2 Candidate Docking score Crash Polarity Control (CNa87) 5.5818 1.3634 3.4231 Preparation Example 1 5.9517 0.8384 3.6834 Preparation Example 6 8.2507 2.1744 3.2779 Preparation Example 7 6.5712 1.3554 2.847 Preparation Example 8 7.8235 2.511 4.4773 Preparation Example 10 5.4865 1.4533 2.798 Preparation Example 11 5.1156 1.011 2.6492

[0091] From the above results, it can be seen that most of the above preparation examples exhibit high binding ability to the AMP-activated protein kinase enzyme as compared with the control CNa87, which makes it possible to predict that such preparation examples can activate the enzyme.

[Experimental Example 2] Measurement of AMP-Activated Protein Kinase Activity

[0092] In order to identify an effect of the Preparation Example 6 having the highest docking score in Experimental Example 1 on activity of the AMP-activated protein kinase (AMPK), a kinase assay was performed. The kinase assay was performed as follows. The control AICAR, which is known to induce activation of the enzyme, and the Preparation Example 6, were administered. Then, purified AMP-activated protein kinase and ATP were allowed to react on a substrate peptide for the AMP-activated protein kinase, and degrees of luminescence thereof were shown as EC.sub.50 in Table 3 and FIG. 3.

[0093] As shown in Table 3 and FIG. 3, the control AICAR exhibited an EC.sub.50 of 0.299, while the Preparation Example 6 exhibited an EC.sub.50 of 0.637 which is about 3-fold higher than the control.

TABLE-US-00003 TABLE 3 Candidate EC.sub.50 (M) Control (AICAR) 0.299 Preparation Example 6 0.637

[0094] From the above results, it can be seen that the Preparation Example 6 according to the present invention not only exhibits high binding ability to the AMP-activated protein kinase, but also increases activity of the enzyme in a very effective manner.

[Preparatory Example 1] Preparation and Breeding of Experimental Animals

[0095] Six-week-old ICR mice (male) were purchased, and then examined for general health for 6 days while subjecting them to quarantine and acclimation under an environment of animal breeding room. Then, healthy individuals were selected and used for the experiment. Identification of each individual was made by marking the number of each individual on the tail using a permanent marker, and identification of a breeding box was made by attaching an individual identification card on which test number, test substance name, test item, date of acquisition, acclimation period, date of grouping, test period, sex/animal number, and person in charge are indicated. After excluding the individuals who developed abnormalities during the acclimation period and the individuals who did not gain weight normally, grouping was carried out so that uniform mean weight and standard deviation are achieved among respective groups.

[0096] A breeding environment for the experimental animals was such that the animals are bred while feeding food and drinking water in an animal breeding room which has been set at a breeding environment of temperature (223 C.), relative humidity (5020) %, the number of ventilation (10 to 15) times/hour, lighting cycle of 12 hours (8:00 to 20:00), illumination (150 to 300) lux, and isolated breeding was carried out during the acclimation and test period.

[Preparatory Example 2] Preparation of Degenerative Neurological Disease-Induced Experimental Animals

[0097] In order to induce a degenerative neurological disease, the experimental animals of Preparatory Example 1 were anesthetized by inhalation with 2.5% isoflurane. Then, 1 g of amyloid-beta peptide.sub.1-42, a degenerative neurological disease-inducing substance, was administered to the animals at both hippocampal positions (AP: 2.3, L: 2.0, H: 1.5) using a stereotaxic apparatus. For the control, 1 g of artificial cerebrospinal fluid made of 145 mM NaCl, 2.7 mM KCl, 1.2 mM CaCl.sub.2, and 1.0 mM MgCl.sub.2 was administered. For the administration, 1 l was injected, at a flow rate of 0.2 l/min, with a 1 ml gas-tight glass syringe (Hamilton, Reno, Nev., USA) using a perfusion pump (Pump 22, Harvard Apparatus, South Natick, Mass., USA). Subsequently, the syringe was allowed to stand for 5 minutes and then removed.

[Experimental Example 3] Morris Water Maze Test

[0098] A behavioral test was conducted to identify whether the compound according to the present invention has a therapeutic effect on degenerative neurological diseases.

[0099] Pre-treatment with donepezil, which is a positive control and a therapeutic agent for degenerative neurological diseases, and the Preparation Example 6 at 1 mg, 10 mg, and 100 mg each was performed. Then, a degenerative neurological disease was induced as in Preparation Example 2. Donepezil and the Preparation Example 6 were administered continuously for 7 days, starting from a degenerative neurological disease-induction day until an experiment end day (anatomy day).

[0100] After induction of the degenerative neurological disease, the following test was conducted after a one-day recovery period passed.

[0101] A circular pool (diameter: 120 cm, height: 75 cm) was filled with water to a depth of 251 cm as in FIG. 4, and a 24 cm platform was prepared by hiding it 1 cm below the water surface. Space cues were installed on the four sides of the wall so that the aminals can identify locations, and skimmed milk powder was dissolved into water so that the animals cannot identify the platform with naked eyes. One day before the experiment, all the experimental animals were allowed to swim freely for 1 minute in a water bath having no platform, so that the animals can adapt to the swimming situation. After that, a test group was allowed to start, in any order, from two of the three quadrants except the quadrant where the platform was placed, and allowed to find the platform for 60 seconds. In a case where an animal finds the platform, the animal was allowed to stay thereon for 20 seconds. In a case where an animal does not find the platform within 45 seconds, the experimenter led the animal to the platform and caused the animal to stay thereon for 20 seconds. As soon as end of the position learning time, which is the 20-second staying time, the animal was allowed to start again from any quadrant, and the time and distance to reach the platform were measured. The procedure was performed four times a day, and all procedures were taken with a camera and analyzed with a video tracking system (Panlab, USA). Probe trials were made on the fifth day after the four-day experiment in total was completed. The test group was allowed to swim freely for 90 seconds without the platform, thereby performing evaluation on whether the test group holds a memory of the platform location. The results are illustrated in FIGS. 5 to 7.

[0102] As illustrated in FIGS. 5 to 7, in a case of the control, on the first day, it found the platform at a level of 95.87 (26.88) seconds in the first trial. Learning progressed as the number of trials and the experimental days passed. In the first trials on respective experimental days, the control found the platform in 95.87 (26.88) seconds, 71.27 (17.83) seconds, 58.93 (22.16) seconds, and 32.37 (8.90) seconds which is on the last fourth day, respectively, and escaped. However, for the beta-amyloid group, which is a negative control, learning did not progress normally. The time taken to find the platform was 143.91 (26.88) seconds at the maximum on the first day, and 60.59 (15.12) seconds at the minimum on the fourth day. From this, it was identified that the escape latency is not decreased normally as compared with the control. The donepezil group found the platform in 111.99 (22.59) seconds at the maximum, and 30.82 (13.96) seconds at the minimum, and escaped.

[0103] On the other hand, for the groups treated with the Preparation Example 6 at 1, 10, and 100 mg, 136.45 (26.02) seconds, 128.59 (23.32) seconds, and 125.66 (19.35) seconds were respectively measured as the maximum platform escape latency. In addition, for the three test groups, 44.45 (13.35) seconds, 40.64 (10.94) seconds, and 36.48 (9.54) seconds were respectively measured as the minimum time to find the platform.

[0104] In addition, in a case where only the first trials on respective experimental days which are indicators showing a long-term memory after 24 hours, are compared among groups, on the first day, no significant difference was observable in all groups except the beta-amyloid group and the control. However, on the second and third days, it was possible to identify that the groups treated with the Preparation Example 6 at 10 mg (second day: 95.0518.06 seconds, p<0.05, third day: 91.6612.68 seconds, p<0.05), 100 mg (second day: 91.6612.68 seconds, p<0.01, third day: 83.4111.46 seconds, p<0.05) exhibit significantly decreased escape latency as compared with the beta-amyloid group (second day: 123.7922.85 seconds, third day: 113.1028.27 seconds). On the last fourth day, groups 1, 10, and 100 mg (1 mg: 73.589.14 seconds, p<0.05, 10 mg: 65.2213.65 seconds, p<0.05, 100 mg: 64.712.63 seconds, p<0.05) exhibited significantly decreased escape latency as compared with the beta-amyloid group (90.3919.77 seconds). In addition, only the last fourth trials, which are indicators showing a short-term memory in repeated trials, were compared among groups. As a result, it was possible to identify that on the first, second, and third days, among the sample groups, 10 mg (first day: 76.1413.27 seconds, p<0.05, second day: 59.8712.56 seconds, p<0.05, third day: 54.0714.38 seconds, p<0.05) and 100 mg (first day: 73.7912.15 seconds, p<0.05, second day: 59.628.22 seconds, p<0.05, third day: 47.7410.81 seconds, p<0.01) exhibit significantly decreased escape latency as compared with the beta-amyloid group (first day: 100.5121.51 seconds, second day: 80.3918.75 seconds, third day: 73.8219.18 seconds). In addition, on the last fourth day, all sample groups (1 mg: 44.4513.35 seconds, p<0.05, 10 mg: 40.6410.94 seconds, p<0.05, 100 mg: 36.489.54 seconds, p<0.01) exhibited significantly decreased escape latency as compared with the beta-amyloid group.

[0105] From the above results, it was found that the Preparation Example 6 recovers memory against degenerative dementia induced by beta-amyloid, thereby decreasing the escape latency.

[Experimental Example 4] Probe Test

[0106] In order to identify whether the compound according to the present invention has a therapeutic effect on degenerative neurological diseases, comprehensive memory ability was tested.

[0107] In the same manner as in Example 3, the Preparation Example 6, the negative control, and donepezil as a positive control were respectively administered, and a probe test was conducted to evaluate comprehensive final memory ability through 4-day repetitive learning. The results are illustrated in FIGS. 8 and 9.

[0108] As illustrated in FIGS. 8 and 9, from the probe test results, it was identified that in a case where the platform is removed, for 90 seconds, the control has the swimming time of 47.19 (6.95) seconds in the quadrant where the platform has been placed, and the beta-amyloid group has the swimming time of 23.90 (6.25) seconds in the quadrant where the platform has been placed, which is significantly decreased as compared with the control. The donepezil group, which is a positive control, had the swimming time of 39.04 (8.68) seconds in the target quadrant, which is significantly increased as compared with the beta-amyloid group (p<0.01). In the test groups, it was identified that all groups (1 mg: 31.086.33 seconds, 10 mg: 36.737.78 seconds, 100 mg: 36.516.55 seconds) exhibit significantly increased swimming time in the target quadrant as compared with the beta-amyloid group. (1 mg: p<0.05, 10, 100 mg: p<0.01).

[Experimental Example 5] Passive Avoidance Response Test

[0109] In the same manner as in Example 3, the Preparation Example 6, the negative control, and donepezil as a positive control were respectively administered. Under a situation where a chamber with a bright light and a dark chamber were connected to each other via a small passage, 4 hours before the experiment, the experimental animals were placed in the chamber with a bright light. The experimental animals instinctively enter the dark chamber. However, after 4 hours of fear learning carried out by using an electric shock to give aversive stimulus, in a case where the experimental animals are placed in the chamber with a bright light, the time until the experimental animals enter the dark chamber was measured and quantified. The results are illustrated in FIG. 10.

[0110] As illustrated in FIG. 10, the above experimental results show that for the control, the time taken to enter the dark chamber starting from the chamber with a bright light was measured as 249.84 (64.84) seconds, and for the beta-amyloid group, the time was 98.21 (43.27) seconds, indicating that the beta-amyloid group exhibits significantly decreased staying time in the chamber with a bright light as compared with the control. In addition, it was identified that among the test groups, groups 10 mg (156.8750.95 seconds) and 100 mg (153.4834.71 seconds) exhibit significantly increased staying time in the chamber with a bright light as compared with the beta-amyloid group (p<0.05).

[Experimental Example 6] Rotarod Test

[0111] In the same manner as in Example 3, the Preparation Example 6, the negative control, and donepezil as a positive control were respectively administered. As an experiment for evaluating effects of the samples on decreased motor coordination ability and motor sensation associated with a degenerative neurological disease, a rotarod experiment which examines the overall motor function of the test groups was conducted. The experimental method was as follows. On the day before the experiment, the animals were subjected to adaptive training by being forced to walk on a rotating (20 rpm) cylinder twice for one minute each time. After 24 hours, the time to lose balance and fall off the rotating cylinder of the same condition was measured, and the result is illustrated in FIG. 11.

[0112] As illustrated in FIG. 11, for the control, the time to keep balance on the cylinder rotating at 15 rpm was 53.62 (13.59) seconds on average, and for the beta-amyloid group, the time was 33.60 (11.95) seconds which is significantly decreased as compared with the control. In the test groups, it was possible to identify that groups 10 mg (48.6210.54 seconds) and 100 mg (47.658.13 seconds) exhibit significantly increased time on the cylinder as compared with the beta-amyloid group (p<0.05).

[Experimental Example 7] Vertical Pole Test

[0113] In the same manner as in Example 3, the Preparation Example 6, the negative control, and donepezil as a positive control were respectively administered. Among effects of the samples on decreased motor coordination ability and motor sensation associated with a degenerative neurological disease, in order to evaluate sensation of balance, experimental animals were placed on the center of a pole inclined at an angle of 45, and then the time taken to lose balance and fall was measured. The results were illustrated in FIG. 12.

[0114] As illustrated in FIG. 12, in a case of the control, the time to keep balance on the pole inclined at an angle of 45 was 14.60 (2.28) seconds on average; and in a case of the beta-amyloid group, the time was significantly decreased to 10.47 (3.50) seconds.

[Experimental Example 8] Measurement of Acetylcholine Hydrolase Activity

[0115] Experimental mice were respectively administered the Preparation Example 6, the negative control, and donepezil as a positive control in the same manner as in Example 3, and the brain tissues were extracted therefrom. Then, acetylcholine hydrolase activity was measured by the following method. To a microplate were continuously added 300 l of 0.1 M Tris buffer, pH 8.0 (Trizma HCl+Trizmabase), 20 l of 0.01 M dithionitrobenzoic acid (DTNB; Sigma, USA), 10 l of brain tissue homogenate (enzyme suspension). Immediately before absorbance measurement, 10 l of 0.1 M acetylthiocholine chloride (Sigma, USA) as a substrate was added thereto. An absorbance meter was used to observe absorbance changes at 405 nm for 5 minutes, so that acetylcholine hydrolase activity (unit/min/mg protein) was measured. The results are illustrated in FIG. 13.

[0116] As illustrated in FIG. 13, the control exhibited an activity level of 0.15 U/mg protein (0.02); however, the beta-amyloid group exhibited activity of 0.31 U/mg protein (0.06) which is a significant difference of about 2.2-fold increase rate. On the other hand, it was identified the donepezil group, which is a positive control, exhibits 0.18 U/mg protein (0.20), indicating significantly decreased acetylcholine hydrolase activity as compared with the beta-amyloid group. Among the sample groups, the groups 10 mg (0.250.07 U/mg protein) and 100 mg (0.240.05 U/mg protein) exhibited significantly decreased enzyme activity.

[Experimental Example 9] Measurement of Acetylcholine Content

[0117] In the same manner as in Example 3, the Preparation Example 6, the negative control, and donepezil as a positive control were respectively administered. Measurement of acetylcholine in brain tissues was performed based on response of o-acyl derivatives with alkaline hydroxylamine. Specifically, 50 l of brain tissue was taken and 50 l of 1% hydroxylamine (Sigma, USA) was added and mixed. Then, pH thereof was adjusted to 1.20.2 using hydrochloric acid. Finally, 500 l of FeCl.sub.3 (10% in 0.1 N HCl) was added to measure an acetylcholine level (umole/mg protein), and then the absorbance was measured at 530 nm. The results are illustrated in FIG. 14.

[0118] As illustrated in FIG. 14, according to the measurement results obtained by the above method, the control showed a content of 25.07 mole/mg protein (1.92). The beta-amyloid group showed a content of 20.48 mole/mg protein (2.05) which is a significant difference and corresponds to a 1.25-fold decrease. However, it was identified that the donepezil group, which is a positive control, shows 23.19 mole/mg protein (1.69), indicating a significantly increased content as compared with the beta-amyloid group. It was identified that the measured values in all sample groups are 1 mg: 22.311.80 mole/mg protein, 10 mg: 23.081.54 mole/mg protein, and 100 mg: 22.791.45 mole/mg protein, indicating a significantly increased content as compared with the beta-amyloid group (1 mg: p<0.05, 10, 100 mg: p<0.01).

[0119] From the above results, it can be seen that the Preparation Example 6 according to the present invention remarkably decreases an acetylcholine content even in a case of being compared with donepezil, a positive control.

[Experimental Example 10] Measurement of Apoptosis Level

[0120] Experimental mice were respectively administered the Preparation Example 6, the negative control, and donepezil as a positive control in the same manner as in Example 3, and the brain tissues were extracted therefrom. Then, an apoptosis level was checked through TUNEL assay. The TUNEL assay is a method to search for cells that have undergone apoptosis by attaching a fluorescent substance to ends of DNA fragments cleaved due to apoptosis. The extracted brain tissue of the experimental animal is fixed by perfusion with 4% formaldehyde. Then, the brain tissue was embedded in paraffin and sectioned to a thickness of 5 um. The brain tissue fixed on a slide was subjected to a TUNEL assay kit so that the end portions where DNA having undergone apoptosis had been fragmented are fluorescence-stained, and checked by a fluorescence microscope. The results are illustrated in FIGS. 15 and 16.

[0121] As illustrated in FIGS. 15 and 16, as a result of conducting the TUNEL assay for all experimental groups in Table 3, the number of Tunel positive cells was observed as 1.1 (1.32) on average in the control; and the number of Tunel positive cells was significantly increased to 115.6 (19.52) in the beta-amyloid group. The donepezil group (113.8314.77), which is a positive control, had no significant change. However, it was identified that all sample groups (1 mg: 88.8318.98, 10 mg: 69.517.87, 100 mg: 67.0019.54) exhibit a significantly decreased number of Tunel positive cells as compared with the beta-amyloid group (1 mg: p<0.05, 10, 100 mg: p<0.01).

[0122] From the above results, it can be seen that the Preparation Example 6 according to the present invention remarkably decreases apoptosis in the brain tissue even in a case of being compared with donepezil, a positive control.

[Experimental Example 11] Physical Property Evaluation: Measurement of Membrane Permeability

[0123] In order to measure a degree of membrane permeability of the Preparation Example 6 according to the present invention, measurement was performed by the PMPA method in which an artificial lipid membrane is formed and permeability is measured by a method other than cells. The results are illustrated in FIG. 17.

[0124] As illustrated in FIG. 17, it was found that the Preparation Example 6 according to the present invention exhibits a membrane permeability measured value of 4.40.152, indicating a medium value in term of degree of permeability.

[0125] From the above results, it can be seen that the Preparation Example 6 according to the present invention has membrane permeability and thus may be indeed effectively utilized clinically.

[Experimental Example 12] Physical Property Evaluation: Measurement of Mutagenicity

[0126] For cancer-causing potential due to the Preparation Example 6 according to the present invention, measurement was performed by the Ames experimental method in which mutagenicity of a test substance is predicted through a mutation reaction test such as substitution, addition, deletion of a few DNA bases using Salmonella typhimurium strains that require a specific amino acid, and thus toxicity thereof is measured.

[0127] In the Ames experimental method, the Preparation Example 6, and 2-nitrofluorene, benzo(a)pyrene, and sodium azide which correspond to the positive controls were respectively dissolved and diluted in DMSO, and treatment with the resultants was performed. Using Salmonella typhimurium strains TA98 and TA100, with or without S9, a degree of bacterial growth depending on the test substance was observed in the control not treated with the Preparation Example 6 and the test group treated therewith. For the Preparation Example 6, a preliminary test with 6-level concentrations obtained by dilution at 1/5 starting from 5000 g/plate, which is the highest treatment concentration in the Ames experimental method, was conducted to select an appropriate concentration that does not exhibit a bacterial growth inhibition phenomenon and toxicity, and a concentration that does not exhibit precipitation during treatment. For each strain, treatment with a single concentration of 200 g/plate was performed in triplicate and mutation was checked. The results are shown in Table 4 below.

TABLE-US-00004 TABLE 4 Con- Reverse mutation/dish Test cen- (mean SD) [factor] strain Compound tration w/o S-9 mix with S-9 mix TA98 Control 0 19 2 23 2 Preparation 200 20 4 35 2 Example 6 [1.1] [1.5] TA100 Control 0 112 16 111 5 Preparation 200 104 12 117 4 Example 6 [0.9] [1.0] TA98 2-Nitrofluorene 2 290 8 Not 915.0] available Benzo(a)pyrene 2 Not 184 17 available [8.0] TA100 Sodium azide 1 618 13 Not [5.5] available Benzo(a)pyrene 2 Not 567 23 available [5.1]

[0128] As shown in Table 4, in the TA98 and TA100 strains, in a case where no metabolic activation enzyme system is applied and in a case where the metabolic activation enzyme system is applied, the concentration groups, in which treatment with the Preparation Example 6 at 200 g/plate each had been performed, did not show a significant increase in reverse mutation as compared with the control. All positive controls exhibited a significantly increased number of colonies as compared with the control. The prepared test substance and the S9 mix were each mixed in the upper agar and culture was performed for 48 hours. As a result, contamination was not observed.

[0129] From the Ames experiment's results which are reliable in view of the fact that the above results show a remarkable difference while showing a 3-fold or higher increase in the number of colonies as compared with the control, it is determined that the Preparation Example 6 is negative since no increase in the number of colonies is observed in a reverse mutation test conducted for bacteria obtained by treating the Salmonella typhimurium TA98 and TA100 strains with the Preparation Example 6. Thus, it can be expected that the Preparation Example 6 according to the present invention is very unlikely to cause cancer.

[Experimental Example 13] Physical Property Test: Cytotoxicity Test

[0130] In order to identify whether the Preparation Example 6 according to the present invention is toxic to cells, a cytotoxicity test was conducted at the Korea Research Institute of Chemical Technology using the Cyto X cell viability assay kit (LPS solution). The results are shown in Table 5 below.

[0131] As shown in Table 5, it can be determined that the Preparation Example 6 has no cytotoxicity, because it can be generally predicted that a compound has no cytotoxicity in a case where IC.sub.50 by the compound is 10 M or higher.

TABLE-US-00005 TABLE 5 IC.sub.50 (M) Compound VERO HFL-1 L929 NIH 3T3 CHO-K1 Preparation 83.6 >100 >100 >100 >100 Example 6

[0132] From the above results, it can be expected that the Preparation Example 6 according to the present invention does not exhibit cytotoxicity in a case of being used in the form of a pharmaceutical composition for preventing or treating degenerative neurological diseases.

[Experimental Example 14] Physical Property Test: Cardiac Stability Test

[0133] Cardiac stability of the Preparation Example 6 according to the present invention was checked using a ligand binding assay. The results are shown in Table 6 below.

[0134] In the ligand binding assay, in a case where an inhibition rate by a 10 M compound is 50 or higher, it is determined that the substance requires attention for binding in hERG channel. As shown in Table 6, it was identified that the Preparation Example 6 exhibits an inhibition rate of 1 or lower per 10 M, and thus would be safe for protein binding in hERG channel.

TABLE-US-00006 TABLE 6 Treatment Concentration (M) Inhibition rate (%) Control 10 69.4 4.78 Preparation Example 10 <1

[0135] From the above results, it can be expected that the Preparation Example 6 according to the present invention has cardiac stability in a case of being used in the form of a pharmaceutical composition for preventing or treating degenerative neurological diseases.

[Experimental Example 15] Organic Physical Property Test

[0136] For a preliminary examination on clinical application of the Preparation Example 6, evaluation on lipophilicity and solubility was performed. The results are shown in Table 7 below.

[0137] As shown in Table 7 below, the Preparation Example 6 was measured to have lipophilicity of 4.75, and solubility of 206.2 and 94.4.

TABLE-US-00007 TABLE 7 Measured item Method Measured value Remark pKa, LogP, ACD/Labs 4.75 lipophilicity T3 (pH-metrior) Solubility Nephelometry 206.2 1.7 M/ DMSO 5%/ 94.4 0.8 g/ml water solvent

[0138] From the above results, it can be seen that the Preparation Example 6 according to the present invention has organic physical properties which make it easy to be used in the form of a pharmaceutical composition for preventing or treating degenerative neurological diseases.

[Experimental Example 16] Physical Property Test: Half-Life

[0139] In order to identify whether the Preparation Example 6 according to the present invention can be orally administered, a half-life of the compound was checked. A request for the half-life test was made to the Korea Research Institute of Chemical Technology, and comparison was made between the half-life of the compound and the half-life of Resveratrol as a control. The results are illustrated in FIG. 18.

[0140] As illustrated in FIG. 18, Resveratrol as a control has a very short half-life of 8 to 10 minutes, while the Preparation Example 6 according to the present invention has a half-life corresponding to 3 hours or longer.

[0141] The above results show that the Preparation Example 6 according to the present invention has a relatively long half-life even in vivo, which enables treatment of degenerative neurological diseases via oral administration.

[0142] Although the present invention has been described in detail above, the scope of the present invention is not limited thereto. It will be obvious to those skilled in the art that various modifications and changes can be made without departing from the technical spirit of the present invention described in the claims.

INDUSTRIAL APPLICABILITY

[0143] A novel compound of the present invention and a pharmaceutical composition comprising the same as an active ingredient induce AMP-activated protein kinase (AMPK) enzyme activation, and thus may be usefully utilized in the fields associated degenerative neurological diseases and the like.