NOVEL ACID SECRETION INHIBITOR AND USE THEREOF

Abstract

The present invention provides a novel compound represented by Chemical Formula 2, or a pharmaceutically acceptable salt thereof. The novel compound according to the present invention exhibits an excellent acid secretion inhibitory effect.

Claims

1. A compound represented by the following Chemical Formula 2 or a pharmaceutically acceptable salt thereof: ##STR00021## in the Chemical Formula 2, X.sub.1 is F; X.sub.2 is hydrogen or F; R.sub.1 is methyl or ethyl; and R.sub.2 is —O(C.sub.1-C.sub.4alkyl) or —(C.sub.1-C.sub.4alkyl).

2. The compound, or the pharmaceutically acceptable salt thereof of claim 1, wherein R.sub.2 is methoxy, ethoxy, methyl or ethyl.

3. The compound, or the pharmaceutically acceptable salt thereof of claim 2, wherein R.sub.2 is methoxy or methyl.

4. The compound, or the pharmaceutically acceptable salt thereof of claim 1, wherein R.sub.1 is methyl.

5. The compound, or the pharmaceutically acceptable salt thereof of claim 1, wherein R.sub.1 is methyl, and R.sub.2 is methoxy or methyl.

6. The compound, or the pharmaceutically acceptable salt thereof of claim 1, wherein X.sub.1 is F; X.sub.2 is F; R.sub.1 is methyl; and R.sub.2 is methoxy or methyl.

7. The compound, or the pharmaceutically acceptable salt thereof of claim 1, wherein X.sub.1 is F; X.sub.2 is hydrogen; R.sub.1 is methyl; and R.sub.2 is methoxy or methyl.

8. The compound, or the pharmaceutically acceptable salt thereof of claim 1, wherein X.sub.1 is F; X.sub.2 is hydrogen or F; R.sub.1 is methyl; and R.sub.2 is methoxy.

9. The compound, or the pharmaceutically acceptable salt thereof of claim 1, wherein X1 is F; X2 is hydrogen or F; R.sub.1 is methyl; and R.sub.2 is methyl.

10. The compound, or the pharmaceutically acceptable salt thereof of claim 1, wherein the compound represented by Chemical Formula 2 is any one selected from the group consisting of the following compounds: 1-(5-(2-fluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine; 1-(5-(2,4-difluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine; 1-(5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine; and 1-(5-(2-fluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine.

11. A pharmaceutical composition comprising the compound according to any one of claims 1 to 10, or the pharmaceutically acceptable salt thereof.

12. A pharmaceutical composition comprising the compound according to any one of claims 1 to 10, or the pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

13. A pharmaceutical composition for preventing or treating gastrointestinal ulcers, gastrointestinal inflammatory diseases, or gastric acid-related diseases, comprising the compound according to any one of claims 1 to 10 or the pharmaceutically acceptable salt thereof.

14. The pharmaceutical composition of claim 13, wherein the gastrointestinal ulcer, gastrointestinal inflammatory disease or gastric acid-related disease is any one or more selected from the group consisting of peptic ulcer, gastric ulcer, duodenal ulcer, NSAID-induced ulcer, acute stress ulcer, Zollinger-Ellison syndrome, Helicobacter pylori infection, gastritis, erosive esophagitis, non-erosive esophagitis, reflux esophagitis, inflammatory bowel disease, symptomatic gastroesophageal reflux disease (symptomatic GERD), functional dyspepsia, gastric cancer, gastric MALT lymphoma, hyperacidity, and upper gastrointestinal hemorrhage due to invasive stress.

15. Use of the compound according to any one of claims 1 to 10, or the pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of gastrointestinal ulcers, gastrointestinal inflammatory diseases or gastric acid-related diseases.

16. The use of claim 15, wherein the gastrointestinal ulcer, gastrointestinal inflammatory disease or gastric acid-related disease is any one or more selected from the group consisting of peptic ulcer, gastric ulcer, duodenal ulcer, NSAID-induced ulcer, acute stress ulcer, Zollinger-Ellison syndrome, Helicobacter pylori infection, gastritis, erosive esophagitis, non-erosive esophagitis, reflux esophagitis, inflammatory bowel disease, symptomatic gastroesophageal reflux disease (symptomatic GERD), functional dyspepsia, gastric cancer, gastric MALT lymphoma, hyperacidity, and upper gastrointestinal hemorrhage due to invasive stress.

17. A method of treating gastrointestinal ulcers, gastrointestinal inflammatory diseases, or gastric acid-related diseases, comprising: administering a therapeutically effective amount of the compound according to any one of claims 1 to 10, or the pharmaceutically acceptable salt thereof to a subject in need thereof.

18. The method of claim 17, wherein the gastrointestinal ulcer, gastrointestinal inflammatory disease or gastric acid-related disease is any one or more selected from the group consisting of peptic ulcer, gastric ulcer, duodenal ulcer, NSAID-induced ulcer, acute stress ulcer, Zollinger-Ellison syndrome, Helicobacter pylori infection, gastritis, erosive esophagitis, non-erosive esophagitis, reflux esophagitis, inflammatory bowel disease, symptomatic gastroesophageal reflux disease (symptomatic GERD), functional dyspepsia, gastric cancer, gastric MALT lymphoma, hyperacidity, and upper gastrointestinal hemorrhage due to invasive stress.

19. A compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of gastrointestinal ulcers, gastrointestinal inflammatory diseases or gastric acid-related diseases.

20. The compound, or the pharmaceutically acceptable salt thereof of claim 19, wherein the gastrointestinal ulcer, gastrointestinal inflammatory disease or gastric acid-related disease is any one or more selected from the group consisting of peptic ulcer, gastric ulcer, duodenal ulcer, NSAID-induced ulcer, acute stress ulcer, Zollinger-Ellison syndrome, Helicobacter pylori infection, gastritis, erosive esophagitis, non-erosive esophagitis, reflux esophagitis, inflammatory bowel disease, symptomatic gastroesophageal reflux disease (symptomatic GERD), functional dyspepsia, gastric cancer, gastric MALT lymphoma, hyperacidity, and upper gastrointestinal hemorrhage due to invasive stress.

Description

Synthetic Example

Synthetic Example 1. Synthesis of Intermediates 1 to 4

[Intermediate 1] methyl 5-(2-fluorophenyl)-4-methoxy-1H-pyrrole-3-carboxylate

Step (1) Synthesis of 2-((tert-butoxycarbonyl)amino)-2-(2-fluorophenyl)acetic acid

[0193] 2-Amino-2-(2-fluorophenyl)acetic acid (1.0 eq., 2.0 g, 11.82 mmol) was dissolved in THF/H.sub.2O=1:1 (70 mL), and then sodium hydrogen carbonate (3.0 eq., 2.98 g, 35.47 mmol) was added, followed by stirring for 30 minutes. Di-tert-butyl dicarbonate (1.2 eq., 3.10 g, 14.18 mmol) was added, and the mixture was stirred at room temperature overnight. THF was removed by reducing the pressure of the reaction solution, and then the pH was adjusted to about 2.5 with 1N HCl aqueous solution. Ethyl acetate (EA) was added, and the resulting mixture was extracted twice. Then, the organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate to obtain 2-((tert-butoxycarbonyl)amino)-2-(2-fluorophenyl)acetic acid as a pale yellow solid (3.0 g, 94%).

Step (2) Synthesis of methyl 4-((tert-butoxycarbonyl)amino)-4-(2-fluorophenyl)-3-oxobutanoate

[0194] 2-((tert-Butoxycarbonyl)amino)-2-(2-fluorophenyl)acetic acid (1.0 eq., 30.0 g, 111.4 mmol) and carbonyldiimidazole (1.03 eq., 18.6 g, 114.7 mmol) were dissolved in acetonitrile (300 mL) and stirred at room temperature for 1 hour. To another flask, monomethyl potassium malonate (1.03 eq., 17.9 g, 114.7 mmol), anhydrous magnesium chloride (1.03 eq., 10.94 g, 114.7 mmol), acetonitrile (300 mL), and triethylamine (1.03 eq., 16 mL, 114.7 mmol) were added and stirred at room temperature for 1 hour. The reactants of the two flasks prepared earlier were mixed using a cannula, and refluxed at 80° C. for 1 hour. After completion of the reaction, the mixture was cooled to room temperature, and water was added thereto. The mixture was cooled with ice, and stirred for 1 hour. The obtained solid was filtered, EA and water were added, and then the pH was adjusted to about 5 using 1N HCl. The mixture was extracted twice with EA, dried, filtered, and concentrated with anhydrous magnesium sulfate to obtain methyl 4-((tert-butoxycarbonyl)amino)-4-(2-fluorophenyl)-3-oxobutanoate as a solid (19.0 g, 52%).

Step (3) Synthesis of 1-(tert-butyl) 3-methyl 5-(fluorophenyl)-4-hydroxy-1H-pyrrole-1,3-dicarboxylate

[0195] Methyl 4-((tert-butoxycarbonyl)amino)-4-(2-fluorophenyl)-3-oxobutanoate (1.0 eq., 15.4 g, 47.33 mmol) and N,N-dimethylformamide dimethyl acetal (3.0 eq., 19 mL, 142.00 mmol) were added to toluene (300 mL) and stirred at 40° C. for 4 hours to complete the reaction. The mixture was evaporated under reduced pressure to remove toluene, and EA and water were added. After neutralization to about pH 7 using 1N HCl, the mixture was extracted twice with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate to obtain 1-(tert-butyl) 3-methyl 5-(fluorophenyl)-4-hydroxy-1H-pyrrole-1,3-dicarboxylate as a solid (14.28 g, 90%).

Step (4) Synthesis of 1-(tert-butyl) 3-methyl 5-(2-fluorophenyl)-4-methoxy-1H-pyrrole-1,3-dicarboxylate

[0196] 1-(tert-Butyl) 3-methyl 5-(fluorophenyl)-4-hydroxy-1H-pyrrole-1,3-dicarboxylate (1.0 eq., 14.28 g, 42.58 mmol), potassium carbonate (2.0 eq., 11.8 g, 85.17 mmol), and dimethyl sulfate (1.13 eq., 4.56 mL, 48.12 mmol) were dissolved in acetone (213 mL) and stirred at 50° C. overnight. The reaction was completed by adding water, and then the excess acetone was removed by evaporation under reduced pressure. After adding EA and water, the mixture was neutralized to about pH 7 using 1N HCl, and then extracted twice with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and purified by silica column chromatography to obtain 1-(tert-butyl) 3-methyl 5-(2-fluorophenyl)-4-methoxy-1H-pyrrole-1,3-dicarboxylate as a solid (14.00 g, 94%).

Step (5) Synthesis of methyl 5-(2-fluorophenyl)-4-methoxy-1H-pyrrole-3-carboxylate (Intermediate 1)

[0197] 1-(tert-Butyl) 3-methyl 5-(2-fluorophenyl)-4-methoxy-1H-pyrrole-1,3-dicarboxylate (1.0 eq., 7.0 g, 20.0 mmol) and trifluoroacetic acid (10.0 eq., 15.3 mL, 200.4 mmol) were dissolved in dichloromethane (35 mL) and stirred at room temperature for 6 hours. After cooling to 0 to 5° C. using ice water, water was added and the pH was adjusted to 7.0 using 50% NaOH aqueous solution. After twice extraction with EA and evaporation, n-hexane was added. Then, the mixture was stirred for 1 hour and filtered to obtain methyl 5-(2-fluorophenyl)-4-methoxy-1H-pyrrole-3-carboxylate obtained as a pale pink solid (4.6 g, 92%).

[0198] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.46 (s, 1H), 7.64 (dt, J=1.6, 7.8 Hz, 1H), 7.36-7.24 (m, 4H), 3.73 (s, 6H).

[Intermediate 2] tert-Butyl ((5-(2-fluorophenyl) methoxy-1H-pyrrol-3-yl)methyl)(methyl)carbamate

Step (1) Synthesis of tert-butyl 2-(2-fluorophenyl)-4-(hydroxymethyl)-3-methoxy-1H-pyrrole-1-carboxylate

[0199] Diisobutylaluminum hydride (1M hexane solution, 5 eq., 64.4 mL, 64.4 mmol) was dissolved in tetrahydrofuran (200 mL), and 1-(tert-butyl) 3-methyl 5-(2-fluorophenyl)-4-methoxy-1H-pyrrole-1,3-dicarboxylate (4.5 g, 12.9 mmol) was slowly added dropwise at 0° C., and stirred at room temperature for 1 hour. Water and a 1N aqueous NaOH solution were sequentially added dropwise, dried over anhydrous magnesium sulfate, filtered through celite, and concentrated. The concentrated residue was purified by column chromatography to obtain tert-butyl 2-(2-fluorophenyl)-4-(hydroxymethyl)-3-methoxy-1H-pyrrole-1-carboxylate as a colorless oil (1.7 g, 41.1%).

[0200] .sup.1H NMR (400 MHz, CDCl.sub.3): δ 7.41-7.33 (m, 2H), 7.30 (s, 1H), 7.19 (dt, J=7.4 Hz, J=1.2 Hz, 1H), 7.10 (dt, J=9.0 Hz, J=0.8 Hz, 1H), 4.61 (d, J=4.8 Hz, 2H), 3.60 (s, 3H), 1.32 (s, 9H)

Step (2) Synthesis of tert-butyl 2-(2-fluorophenyl)-4-formyl-3-methoxy-1H-pyrrole-1-carboxylate

[0201] tert-Butyl 2-(2-fluorophenyl)-4-(hydroxymethyl) methoxy-1H-pyrrole-1-carboxylate (1.7 g, 5.3 mmol) was dissolved in dichloromethane (20 mL), and Dess-Martin periodinane (1 eq., 2.24 g, 5.3 mmol) was slowly added dropwise and stirred at room temperature for 1 hour. Celite was added to the reaction mixture. The resulting product was concentrated and purified by column chromatography to obtain (5-(2-fluorophenyl)-4-methoxy-1H-pyrrol-3-yl)methanol as a colorless oil (1.23 g, 72.8%).

[0202] .sup.1H NMR (400 MHz, CDCl.sub.3): δ 9.89 (s, 1H), 7.92 (s, 1H), 7.42-7.37 (m, 2H), 7.22 (dt, J=7.5 Hz, J=0.9 Hz, 1H), 7.12 (dt, J=9.2 Hz, J=0.9 Hz, 1H), 3.75 (s, 3H), 1.38 (s, 9H)

Step (3) Synthesis of 5-(2-fluorophenyl)-4-methoxy-1H-pyrrole-3-carbaldehyde

[0203] tert-Butyl 2-(2-fluorophenyl)-4-formyl-3-methoxy-1H-pyrrole-1-carboxylate (1.2 g, 3.8 mmol) was dissolved in water/methanol (1/3, 20 mL), potassium carbonate (3 eq., 1.6 g, 11.3 mmol) was added dropwise, and then the mixture was stirred at 100° C. for 2.5 hours. The reaction product was dried over anhydrous sodium sulfate and filtered to obtain 5-(2-fluorophenyl)-4-methoxy-1H-pyrrole-3-carbaldehyde as a yellow solid (800.0 mg, 97.1%).

[0204] .sup.1H NMR (400 MHz, CDCl.sub.3): δ 9.87 (s, 1H), 9.11 (brs, 1H), 8.17-8.13 (m, 1H), 7.34 (d, J=4.0 Hz, 1H), 7.27-7.23 (m, 2H), 7.19-7.16 (m, 1H), 3.98 (s, 3H)

Step (4) Synthesis of tert-butyl ((5-(2-fluorophenyl)-4-methoxy-1H-pyrrol-3-yl)methyl)(methyl)carbamate (Intermediate 2)

[0205] 5-(2-Fluorophenyl)-4-methoxy-1H-pyrrole carbaldehyde (800 mg, 3.65 mmol) was dissolved in methanol (50 mL), 40% methylamine solution (2.3 eq., 0.86 mL, 8.4 mmol) was added dropwise, and the mixture was stirred at room temperature for 30 minutes. The reaction mixture was cooled to 0° C., sodium borohydride (1.5 eq., 207.1 mg, 5.5 mmol) was added dropwise, and then the mixture was stirred at room temperature for 30 minutes. Water (150 mL) was added dropwise to the reaction mixture, the resulting product was stirred for 1 hour at the same temperature, and brine was added dropwise, followed by extraction with EA. The extracted organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate. The concentrated residue was dissolved in acetonitrile (40 mL), and then di-tert-butyl dicarbonate (1.2 eq., 955.7 mg, 4.4 mmol) was added dropwise, followed by stirring at room temperature for 2 hours. Water and EA were added dropwise to the reaction mixture for extraction, and the organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate. The concentrated residue was purified by column chromatography to obtain tert-butyl ((5-(2-fluorophenyl)-4-methoxy-1H-pyrrol yl)methyl) (methyl)carbamate as a light brown solid (946.8 mg, 78.9%).

[0206] .sup.1H NMR (400 MHz, CDCl.sub.3): δ 8.55 (brs, 1H), 8.07 (dt, J=7.9 Hz, J=1.7 Hz, 1H), 7.20-7.06 (m, 1H), 6.62 (s, 1H), 4.35 (s, 2H), 3.72 (s, 3H), 2.86 (s, 3H), 1.50 (s, 9H)

[Intermediate 3] Methyl 5-(2,4-difluorophenyl)-4-methoxy-1H-pyrrole-3-carboxylate

Step (1) Synthesis of 2-((tert-butoxycarbonyl)amino)-2-(2,4-difluorophenyl)acetic acid

[0207] 2-Amino-2-(2,4-difluorophenyl)acetic acid (1.0 eq., 7.22 g, 38.6 mmol) was dissolved in THF/H.sub.2O (1:1, 200 mL), and then cooled to 0° C. NaHCO.sub.3 (3.0 eq., 9.74 g, 116 mmol) and Boc.sub.2O (1.2 eq., 10.64 mL, 46.3 mmol) were added, and after stirring at room temperature overnight, water was added to the reaction solution, and the pH was adjusted to 2.5. Then, the resulting mixture was extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate to obtain 2-((tert-butoxycarbonyl)amino)-2-(2,4-difluorophenyl)acetic acid (17.35 g, 99%) as a white solid without further purification. [M+Na]+: 310

Step (2) Synthesis of methyl 4-((tert-butoxycarbonyl)amino)-4-(2,4-difluorophenyl)-3-oxobutanoate

[0208] 2-((tert-Butoxycarbonyl)amino)-2-(2,4-difluorophenyl)acetic acid (1.0 eq., 38.6 mmol) and carbonyldiimidazole (1.1 eq., 6.89 g, 42.5 mmol) were dissolved in acetonitrile (100 mL). In another flask, methyl potassium malonate (1.1 eq., 6.64 g, 42.5 mmol), triethylamine (1.1 eq., 5.97 mL, 42.5 mmol), magnesium chloride (1.1 eq., 4.05 g, 42.5 mmol) were dissolved in acetonitrile. (100 mL). After each solution was stirred at room temperature for 1 hour, the two solutions prepared above were combined and stirred at 80° C. for 3 hours. After adding H.sub.2O (100 mL), the mixture was stirred at room temperature for 2 hours, and the resulting solid was filtered. EA and water were added to the filtered solid, and the mixture was stirred at room temperature for 10 minutes and neutralized to pH 7 with aq. HCl. The organic layer extracted with EA was dried, filtered, and concentrated with anhydrous magnesium sulfate, concentrated to obtain methyl 4-((tert-butoxycarbonyl)amino)-4-(2,4-difluorophenyl)-3-oxobutanoate (12.64 g, 95%) as a brown liquid without further purification.

[0209] .sup.1H NMR (400 MHz, CDCl.sub.3): δ 7.33-7.28 (m, 1H), 6.97-6.87 (m, 2H), 5.85 (brs, 1H), 5.67 (d, J=6.8 Hz, 1H), 3.70 (s, 3H), 3.53 (d, J=16.0 Hz, 1H), 4.75 (d, J=16.0 Hz, 1H), 1.34 (s, 9H).

Step (3) Synthesis of 1-(tert-butyl) 3-methyl 5-(2,4-difluorophenyl)-4-hydroxy-1H-pyrrole-1,3-dicarboxylate

[0210] Methyl 4-((tert-butoxycarbonyl)amino)-4-(2,4-difluorophenyl)-3-oxobutanoate (1.0 eq., 12.64 g, 36.8 mmol) and N,N-dimethylformamide dimethyl acetal (3 eq., 14.7 mL, 110.4 mmol) were dissolved in toluene (184 mL) and stirred at 40° C. for 5 hours. After concentration, EA and water were added, and the mixture was neutralized to pH 7 with 1N HCl. The organic layer extracted with EA was dried, filtered, and concentrated with anhydrous magnesium sulfate to obtain 1-(tert-butyl) 3-methyl 5-(2,4-difluorophenyl)-4-hydroxy-1H-pyrrole-1,3-dicarboxylate as a brown liquid without further purification.

[0211] .sup.1H NMR (400 MHz, CDCl.sub.3): δ 7.75 (s, 1H), 7.52 (s, 1H), 7.43 (dt, J=8.4, 6.8 Hz, 1H), 6.98-6.84 (m, 2H), 3.92 (s, 3H), 1.41 (s, 9H).

Step (4) Synthesis of 1-(tert-butyl) 3-methyl 5-(2,4-difluorophenyl)-4-methoxy-1H-pyrrole-1,3-dicarboxylate

[0212] 1-(tert-Butyl) 3-methyl 5-(2,4-difluorophenyl)-4-hydroxy-1H-pyrrole-1,3-dicarboxylate (1.0 eq., 36.8 mmol), potassium carbonate (2.0 eq., 10.2 g, 73.6 mmol), and dimethyl sulfate (1.2 eq., 4.2 mL, 44.2 mmol) were dissolved in acetone (184 mL), and stirred at 40° C. overnight. EA and water were added, and the mixture was neutralized to pH 7 with 1N HCl. The organic layer extracted with EA was dried, filtered, and concentrated with anhydrous magnesium sulfate, purified by silica chromatography to obtain 1-(tert-butyl) 3-methyl 5-(2,4-difluorophenyl)-4-methoxy-1H-pyrrole-1,3-dicarboxylate (12.86 g, 95%) as a yellow liquid. [M+H]+: 367

[0213] .sup.1H NMR (400 MHz, CDCl.sub.3): δ 7.90 (s, 1H), 7.36 (dt, J=8.4, 6.4 Hz, 1H), 6.99-6.86 (m, 2H), 3.90 (s, 3H), 3.70 (s, 3H), 1.41 (s, 9H).

Step (5) Synthesis of methyl 5-(2,4-difluorophenyl) methoxy-1H-pyrrole-3-carboxylate (Intermediate 3)

[0214] 1-(tert-Butyl) 3-methyl 5-(2,4-fluorophenyl)-4-methoxy-1H-pyrrole-1,3-dicarboxylate (1.0 eq., 12.8 g, 34.8 mmol) and trifluoroacetic acid (10 eq., 26 mL, 348 mmol) were dissolved in dichloromethane (70 mL) and stirred at room temperature for 5 hours. Water (100 mL) was added at 0° C. and the mixture was neutralized to pH 7 with 1N NaOH. The reaction solution was extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and solidified with hexane and EA to obtain methyl 5-(2,4-difluorophenyl)-4-methoxy-1H-pyrrole-3-carboxylate as a pale pink solid (4.98 g, 54%) without further purification.

[0215] .sup.1H NMR (400 MHz, CDCl.sub.3): δ 8.82 (brs, 1H), 8.14 (dt, J=9.1, 6.5 Hz, 1H), 7.33 (d, J=3.6 Hz, 1H), 7.01-6.87 (m, 2H), 3.89 (s, 3H), 3.88 (s, 3H).

[Intermediate 4] tert-Butyl ((5-(2,4-difluorophenyl)-4-methoxy-1H-pyrrol-3-yl)methyl)(methyl)carbamate

Step (1) Synthesis of tert-butyl 2-(2,4-difluorophenyl)-4-(hydroxymethyl)-3-methoxy-1H-pyrrole-1-carboxylate

[0216] 1-(tert-Butyl) 3-methyl 5-(2,4-difluorophenyl) methoxy-1H-pyrrole-1,3-dicarboxylate (1.0 eq., 10.02 g, 27.3 mmol) was dissolved in THF (137 mL) and cooled to 0° C. 1.0 M DIBAL-H (8.0 eq., 219 mL, 219 mmol) was slowly added in THF. The reaction solution was stirred at room temperature for 2 hours. After cooling to 0° C., H.sub.2O (8.76 mL), 15% NaOH (8.76 mL), and H.sub.2O (22 mL) were sequentially added. Then, after stirring at room temperature for 20 minutes, anhydrous magnesium sulfate was added, and the mixture was stirred for 20 minutes and filtered through Celite. After concentration, purification was performed by silica chromatography to obtain tert-butyl 2-(2,4-difluorophenyl)-4-(hydroxymethyl)-3-methoxy-1H-pyrrole-1-carboxylate (5.13 g, 55%).

[0217] .sup.1H NMR (400 MHz, CDCl.sub.3): δ 7.35 (dt, J=8.4, 6.4 Hz, 1H), 7.29 (s, 1H), 6.97-6.85 (m, 2H), 4.6 (s, 2H), 3.60 (s, 3H), 1.37 (s, 9H).

Step (2) Synthesis of tert-butyl 2-(2,4-difluorophenyl)-4-formyl-3-methoxy-1H-pyrrole-1-carboxylate

[0218] tert-Butyl 2-(2,4-difluorophenyl)-4-(hydroxymethyl)-3-methoxy-1H-pyrrole-1-carboxylate (1.0 eq., 5.13 g, 15.1 mmol) was dissolved in dichloromethane (76 mL) and cooled to 0° C. DMP (1.1 eq., 7.04 g, 16.6 mmol) was added, followed by stirring at room temperature for 30 minutes. The reaction solution was washed with aq. NaOH and then extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and purified by silica chromatography to obtain tert-butyl 2-(2,4-difluorophenyl)-4-formyl-3-methoxy-1H-pyrrole-1-carboxylate (3.56 g, 70%) as a yellow solid.

[0219] .sup.1H NMR (400 MHz, CDCl.sub.3): δ 9.88 (s, 1H), 7.91 (s, 1H), 7.36 (dt, J=8.4, 6.4 Hz, 1H), 7.00-6.90 (m, 2H), 3.75 (s, 3H), 1.41 (s, 9H).

Step (3) Synthesis of 5-(2,4-difluorophenyl)-4-methoxy-1H-pyrrole-3-carbaldehyde

[0220] tert-Butyl 2-(2,4-difluorophenyl)-4-formyl-3-methoxy-1H-pyrrole-1-carboxylate (1.0 eq., 3.56 g, 10.6 mmol) and potassium carbonate (3 eq., 4.40 g, 31.8 mmol) were dissolved in methanol/H.sub.2O (3:1, 104 mL), and stirred at 110° C. for 1 hour. After concentration, the concentrated product was filtered with acetone, and solidified with dichloromethane and hexane to obtain 5-(2,4-difluorophenyl)-4-methoxy-1H-pyrrole-3-carboaldehyde (2.17 g, 86%) as an orange solid without further purification.

[0221] .sup.1H NMR (400 MHz, CDCl.sub.3): δ 9.86 (s, 1H), 8.97 (brs, 1H), 8.12 (dt, J=9.0, 6.4 Hz, 1H), 7.33 (d, J=4.0 Hz, 1H), 7.03-6.91 (m, 2H), 3.96 (s, 3H).

Step (4) Synthesis of tert-butyl ((5-(2,4-difluorophenyl)-4-methoxy-1H-pyrrol-3-yl)methyl)(methyl)carbamate (Intermediate 4)

[0222] 2.0 M methyl amine was dissolved in methanol (90 mL), followed by stirring at room temperature for 30 minutes in 5-(2,4-Difluorophenyl)-4-methoxy-1H-pyrrole-3-carboaldehyde (1.0 eq., 2.17 g, 9.15 mmol), THF (10 eq., 46 mL, 91.5 mmol). NaBH.sub.4 (5 eq., 1.73 g, 45.8 mmol) was added and the mixture was stirred at room temperature for 30 minutes. Then, water was added and the mixture was stirred for an additional 30 minutes. The reaction solution was washed with brine and extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate. Then, the concentrated product was dissolved immediately in acetonitrile (46 mL), Boc.sub.2O (1.2 eq., 2.53 mL, 11.0 mmol) was slowly added thereto, and the resulting mixture was stirred at room temperature for 1 hour. Water was added to the reaction solution, followed by extraction with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and purified by silica chromatography to obtain tert-butyl 5-(2,4-difluorophenyl)-4-methoxy-1H-pyrrol-3-yl)methyl) (methyl)carbamate (2.46 g, 76%) as a brown solid.

[0223] .sup.1H NMR (400 MHz, CDCl.sub.3): δ 8.45 (brs, 1H), 8.05 (dt, J=9.0, 6.5 Hz, 1H), 6.98-6.85 (m, 2H), 6.63 (brs, 1H), 4.37 (brs, 2H), 3.73 (s, 3H), 2.88 (s, 3H), 1.52 (s, 9H).

[0224] Compounds of Examples 1 to 6 below were synthesized using the synthesized intermediates 1 to 4. The synthesis methods thereof are based on Reaction Schemes 1 and 2 above. As an example of the preparation for the Example compounds above, the preparation methods of Examples 1 to 6 below were specifically described.

[0225] Hereinafter, the synthesis methods of Examples 1 to 6 are shown in detail.

Synthesis Example 1. Synthesis of Example 1

[Example 1] 1-(5-(2-Fluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine

Step (1) Synthesis of methyl 5-(2-fluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrole-3-carboxylate

[0226] Methyl 5-(2-fluorophenyl)-4-methoxy-1H-pyrrole-3-carboxylate (intermediate 1, 1.0 eq., 1.2 g, 4.8 mmol) was dissolved in THF (20.0 mL), and NaH (2.0 eq., 384.8 mg, 9.6 mmol) was added dropwise at 0° C., followed by stirring at room temperature for 10 minutes. 6-Methoxypyridine-3-sulfonyl chloride (1.5 eq., 1.6 g, 7.2 mmol) was added, followed by stirring at room temperature for 1 hour. Water was added to the reaction solution, and the resulting solution was extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and purified by column chromatography to obtain methyl 5-(2-fluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrole-3-carboxylate as a light brown solid (1.85 g, 91.6%).

Step (2) Synthesis of 5-(2-fluorophenyl)-4-methoxy ((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)methanol

[0227] Methyl 5-(2-fluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrole-3-carboxylate (1.0 eq., 1.0 g, 2.38 mmol) was dissolved in THF (5.0 mL), and DIBAL 1.0 M was added dropwise in n-hexane solution (5.0 eq., 11.9 mL, 11.9 mmol) at 0° C., followed by stirring at room temperature for 1 hour. The reaction solution was cooled to 0° C., the reaction was completed with an aqueous Rochelle salt solution, and the resulting solution was extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and purified by column chromatography to obtain 5-(2-fluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)methanol as a yellow oil (654.8 mg, 70.2%).

Step (3) Synthesis of methyl 5-(2-fluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrole-3-carbaldehyde

[0228] 5-(2-Fluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)methanol (1.0 eq., 500.0 mg, 1.3 mmol) and Dess-Martin periodinane (1.0 eq., 540.4 mg, 1.3 mmol) were dissolved in DCM (10.0 mL) and stirred at room temperature for 1 hour. The reaction product was concentrated and purified by column chromatography to obtain 5-(2-fluorophenyl)-4-methoxy-1-((6-methoxypyridin yl)sulfonyl)-1H-pyrrole-3-carbaldehyde as a pale dark blue solid (388.2 mg, 78.1%).

Step (4) Synthesis of 1-(5-(2-Fluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine

[0229] 5-(2-Fluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrole-3-carbaldehyde (1.0 eq., 385.0 mg, 0.99 mmol) was dissolved in THF (5.0 mL), and 2.0 M of methylamine was added in THF (10 eq., 4.9 mL, 9.9 mmol). After stirring at room temperature for 1 hour, the reaction product was cooled to 0° C., NaBH.sub.4 (10 eq., 373.4 mg, 9.9 mmol) was added, and the resulting mixture was stirred at room temperature for 1 hour. To the reaction solution, 6.0N aqueous hydrogen chloride solution was slowly added dropwise, and the resulting solid was filtered. The filtered solid was dissolved in water, and 1N aqueous sodium hydroxide solution was added thereto, followed by extraction with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate to obtain 1-(5-(2-fluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine as a white solid (125.8 mg, 28.3%) [M+H].sup.+: 405.

Synthesis Example 2. Synthesis of Example 2

[Example 2] 1-(5-(2,4-Difluorophenyl)-4-methoxy ((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine

Step (1) Synthesis of methyl 5-(2,4-difluorophenyl) methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrole carboxylate

[0230] Methyl 5-(2,4-difluorophenyl)-4-methoxy-1H-pyrrole-3-carboxylate (intermediate 3, 1.0 eq., 802 mg, 3.00 mmol) and NaH (1.5 eq., 180 mg, 4.5 mmol) were dissolved in anhydrous DMF (15.0 mL) and stirred at room temperature for 10 minutes. 6-Methoxypyridine-3-sulfonyl chloride (1.5 eq., 934 mg, 4.50 mmol) was added, followed by stirring at room temperature for 1 hour. After adding distilled water to the reaction solution, the resulting solution was washed with brine and extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and purified by column chromatography to obtain methyl 5-(2,4-difluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrole-3-carboxylate (1.09 g, 83%).

Step (2) Synthesis of (5-(2,4-difluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)methanol

[0231] Methyl 5-(2,4-difluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrole-3-carboxylate (1.0 eq., 1.09 g, 2.49 mmol) was dissolved in anhydrous THF (13.0 mL), and then DIBAL 1.0 M was added dropwise in THF (5.0 eq., 12.4 mL, 12.4 mmol) at 0° C. Then, the reaction solution was stirred at room temperature for 4 hours. To the reaction solution, 0.50 mL of water, 0.5 mL of 1N aqueous sodium hydroxide solution, and 1.25 mL of water were sequentially added. Then, the resulting mixture was stirred for 30 minutes, and anhydrous magnesium sulfate was added, followed by stirring for 30 minutes. The resulting product was dried, filtered, concentrated, and purified by column chromatography to obtain (5-(2,4-difluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)methanol (869 mg, 85%).

Step (3) Synthesis of 5-(2,4-difluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrole-3-carbaldehyde

[0232] 5-(2,4-Difluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl) sulfonyl)-1H-pyrrol-3-yl)methanol (1.0 eq., 869 mg, 2.12 mmol) and Dess-Martin periodinane (1.1 eq., 988 mg, 2.33 mmol) were dissolved in DCM (21.0 mL) and stirred at room temperature for 30 minutes. NaHCO.sub.3 aqueous solution was added to the reaction solution, and the resulting solution was washed with Na.sub.2S.sub.2O.sub.3 aqueous solution and extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and purified by column chromatography to obtain 5-(2,4-difluorophenyl)-4-methoxy-1-((6-methoxypyridin yl)sulfonyl)-1H-pyrrole-3-carbaldehyde (824 mg, 95%).

Step (4) Synthesis of 1-(5-(2,4-difluorophenyl) methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine

[0233] 5-(2,4-Difluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrole-3-carbaldehyde (1.0 eq., 824 mg, 2.02 mmol) was dissolved in THF (20.0 mL), and 2.0M of methylamine was added in THF (20 eq., 20.2 mL, 40.4 mmol). After stirring at room temperature for 3 hours, NaBH.sub.4 (10 eq., 764 mg, 20.2 mmol) was added, followed by stirring for 18 hours. After adding distilled water to the reaction solution, the resulting solution was washed with brine and extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and purified by column chromatography to obtain 1-(5-(2,4-difluorophenyl)-4-methoxy-1-((6-methoxypyridin-3-yl)sulfonyl)-1H-pyrrole-3-yl)-N-methylmethanamine as a red syrup (90.0 mg, 10%) [M+H].sup.+: 423.

Synthesis Example 3. Synthesis of Example 3

[Example 3] 1-(5-(2,4-Difluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine

Step (1) Synthesis of methyl 5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrole-3-carboxylate

[0234] Methyl 5-(2,4-difluorophenyl)-4-methoxy-1H-pyrrole carboxylate (intermediate 3, 1.0 eq., 534 mg, 2.0 mmol) and NaH (1.5 eq., 120 mg, 3.0 mmol) were dissolved in anhydrous DMF (10.0 mL) and stirred at 50° C. for 50 minutes. 6-Methylpyridine-3-sulfonyl chloride (1.5 eq., 575 mg, 3.0 mmol) was added, followed by stirring at 50° C. for 16 hours. After adding distilled water to the reaction solution, the resulting solution was washed with brine and extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and purified by column chromatography to obtain methyl 5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrole-3-carboxylate (614 mg, 73%).

Step (2) Synthesis of (5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)methanol

[0235] Methyl 5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrole-3-carboxylate (1.0 eq., 614 mg, 1.45 mmol) was dissolved in anhydrous THF (7.27 mL), and then DIBAL 1.0M was added dropwise in THF (5.0 eq., 7.27 mL, 7.27 mmol) at 0° C. Then, the resulting mixture was stirred at room temperature for 30 minutes. To the reaction solution, 0.29 mL of water, 0.29 mL of 15% aqueous sodium hydroxide solution, and 0.73 mL of water were sequentially added. Then, the resulting mixture was stirred for 14 hours. Anhydrous magnesium sulfate was added, followed by stirring for 30 minutes. The resulting product was dried, filtered, concentrated, and purified by column chromatography to obtain (5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)methanol as a yellow solid (494 mg, 86%).

Step (3) Synthesis of 5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrole-3-carbaldehyde

[0236] 5-(2,4-Difluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)methanol (1.0 eq., 494 mg, 1.25 mmol) and Dess-Martin periodinane (1.1 eq., 583 mg, 1.38 mmol) were dissolved in DCM (12.0 mL) and stirred at room temperature for 40 minutes. An aqueous sodium hydroxide solution was added to the reaction solution, followed by extraction with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and purified by column chromatography to obtain 5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrole-3-carbaldehyde (422 mg, 86%).

Step (4) Synthesis of 1-(5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine

[0237] 5-(2,4-Difluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrole-3-carbaldehyde (1.0 eq., 422 mg, 1.07 mmol) was dissolved in MeOH (5.0 mL), and 2.0M of methylamine was added in THF (10 eq., 5.2 mL, 10.7 mmol). After stirring at room temperature for 30 minutes, NaBH.sub.4 (5 eq., 204 mg, 5.38 mmol) was added, followed by stirring for 10 minutes. After adding NaHCO.sub.3 aqueous solution to the reaction solution, the resulting solution was washed with brine and extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and purified by column chromatography to obtain 1-(5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrole-3-yl)-N-methylmethanamine as a yellow solid (176 mg, 40%) [M+H]+: 408.

Synthesis Example 4. Synthesis of Example 4

[Example 4] 1-(5-(2-Fluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine

Step (1) Synthesis of tert-butyl ((5-(2-fluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate

[0238] tert-Butyl ((5-(2-fluorophenyl)-4-methoxy-1H-pyrrol-3-yl)methyl) (methyl)carbamate (intermediate 2, 100.0 mg, 0.3 mmol), NaH (24.0 mg, 0.6 mmol), and 15-crown-5-ether (0.9 mL, 0.5 mmol) were dissolved in anhydrous THF (1.5 mL) and stirred at 50° C. for 10 minutes. 6-Methylpyridine-3-sulfonyl chloride (86.0 mg, 0.5 mmol) was added and stirred at room temperature for 30 minutes. After adding distilled water to the reaction solution, the resulting solution was washed with brine and extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and purified by column chromatography to obtain tert-butyl ((5-(2-fluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)methyl) (methyl)carbamate as a pale yellow oil (60.9 mg, 42%).

Step (2) Synthesis of 1-(5-(2-fluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine

[0239] tert-Butyl ((5-(2-fluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrol-3-yl)methyl) (methyl)carbamate (60.0 mg, 0.1 mmol), and 1.0 M hydrogen chloride in an ethyl acetate solution (2.0 mL) were dissolved in ethanol (1.0 mL) and stirred at room temperature for 4 hours. After adding NaHCO.sub.3 aqueous solution to the reaction solution, the resulting solution was washed with brine and extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and purified by column chromatography to obtain 1-(5-(2-fluorophenyl)-4-methoxy-1-((6-methylpyridin-3-yl)sulfonyl)-1H-pyrrole-3-yl)-N-methylmethanamine as a light yellow solid (49.8 mg, 62%) [M+H].sup.+: 390.

Synthesis Example 5. Example 5

[Example 5] 1-5-(2,4-Difluorophenyl)-4-methoxy-1-((6-methylpyridin-2-yl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine

Step (1) Synthesis of methyl 5-(2,4-difluorophenyl) methoxy-1-((6-methylpyridin-2-yl)sulfonyl)-1H-pyrrole-3-carboxylate

[0240] Methyl 5-(2,4-difluorophenyl)-4-methoxy-1H-pyrrole-3-carboxylate (intermediate 3, 1.0 eq., 400.0 mg, 1.5 mmol) and NaH (1.5 eq., 90.0 mg, 2.25 mmol) were dissolved in anhydrous DMF (10.0 mL) and stirred at room temperature for 30 minutes. 6-Methylpyridine-2-sulfonyl chloride (1.5 eq., 430 mg, 2.25 mmol) was added, followed by stirring at room temperature for 5 hours. After adding distilled water to the reaction solution, the resulting solution was washed with brine and extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and purified by column chromatography to obtain methyl 5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-2-yl)sulfonyl)-1H-pyrrole-3-carboxylate as a clear syrup (442.0 mg, 70%).

Step (2) Synthesis of (5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-2-yl)sulfonyl)-1H-pyrrol-3-yl)methanol

[0241] Methyl 5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-2-yl)sulfonyl)-1H-pyrrole-3-carboxylate (1.0 eq., 439.0 mg, 1.04 mmol) was dissolved in anhydrous THF (5.0 mL), and then DIBAL 1.0M was added dropwise in THF (3.0 eq., 3.12 mL, 3.12 mmol) at 0° C. Then, the reaction solution was stirred at room temperature for 2 hours. MeOH was added to the reaction solution, and then the resulting solution was washed with an aqueous Rochelle salt solution and extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate to obtain (5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-2-yl)sulfonyl)-1H-pyrrol-3-yl)methanol as yellow syrup (417.0 mg, 102%).

Step (3) Synthesis of 5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-2-yl)sulfonyl)-1H-pyrrole-3-carbaldehyde

[0242] 5-(2,4-Difluorophenyl)-4-methoxy-1-((6-methylpyridin-2-yl)sulfonyl)-1H-pyrrol-3-yl)methanol (1.0 eq., 398.0 mg, 1.01 mmol) and Dess-Martin periodinane (1.0 eq., 428.0 mg, 1.01 mmol) were dissolved in DCM (10.0 mL) and stirred at room temperature for 5 hours. NaHCO.sub.3 aqueous solution was added to the reaction solution, and the resulting solution was washed with Na.sub.2S.sub.2O.sub.3 aqueous solution and extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and purified by column chromatography to obtain 5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-2-yl)sulfonyl)-1H-pyrrole carbaldehyde as a yellow syrup (331.0 mg, 84%).

Step (4) Synthesis of 1-5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-2-yl)sulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine

[0243] 5-(2,4-Difluorophenyl)-4-methoxy-1-((6-methylpyridin-2-yl)sulfonyl)-1H-pyrrole-3-carbaldehyde (1.0 eq., 331.0 mg, 0.84 mmol) was dissolved in MeOH (8.5 mL), and 9.8M of methylamine was added in MeOH (20 eq., 1.72 mL, 16.9 mmol). After stirring at room temperature for 1 hour, NaBH4 (10 eq., 318.0 mg, 8.4 mmol) was added, followed by stirring for 30 minutes. After adding NaHCO.sub.3 aqueous solution to the reaction solution, the resulting solution was washed with brine and extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and purified by column chromatography to obtain 1-5-(2,4-difluorophenyl)-4-methoxy-1-((6-methylpyridin-2-yl)sulfonyl)-1H-pyrrole-3-yl)-N-methylmethanamine as a yellow syrup (185.0 mg, 54%) [M+H]+: 407.

Synthesis Example 6. Synthesis of Example 6

[0244] [Example 6] 1-(5-(2-Fluorophenyl)-4-methoxy-1-(pyridin-2-ylsulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine

Step (1) Synthesis of tert-butyl ((5-(2-fluorophenyl)-4-methoxy-1-(pyridin-2-ylsulfonyl)-1H-pyrrol-3-yl)methyl)(methyl)carbamate

[0245] tert-Butyl ((5-(2-fluorophenyl)-4-methoxy-1H-pyrrol-3-yl)methyl) (methyl)carbamate (intermediate 2, 1.0 eq.), NaH (1.5 eq., 90.0 mg, 2.25 mmol), and 15-crown-5-ether (catalytic amount) were dissolved in anhydrous THF (10.0 mL) and stirred at room temperature for 30 minutes. Pyridine-2-sulfonyl chloride (1.5 eq., 430 mg, 2.25 mmol) was added, followed by stirring at room temperature for 5 hours. After adding distilled water to the reaction solution, the resulting solution was washed with brine and extracted with EA. The organic layer was dried, filtered, and concentrated with anhydrous magnesium sulfate, and purified by column chromatography to obtain tert-butyl ((5-(2-fluorophenyl)-4-methoxy-1-(pyridin-2-ylsulfonyl)-1H-pyrrol-3-yl)methyl) (methyl)carbamate as a brown oil (80 mg, 55%).

Step (2) Synthesis of 1-(5-(2-Fluorophenyl)-4-methoxy-1-(pyridin-2-ylsulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine

[0246] tert-Butyl ((5-(2-fluorophenyl)-4-methoxy-1-(pyridin-2-ylsulfonyl)-1H-pyrrol-3-yl)methyl) (methyl)carbamate (0.17 eq., 80 mg) and trifluoroacetic acid (10.0 eq., 0.88 mL, 11.54 mmol) were dissolved in dichloromethane (2.3 mL) and stirred at room temperature for 6 hours. After removing the solvent by distillation under reduced pressure, the resulting product was cooled to 0 to 5° C. using ice water, then water was added and the pH was adjusted to 7.0 using an aqueous NaHCO.sub.3 solution. After twice extraction with EA and evaporation, n-hexane was added, and the resulting product was stirred for 1 hour and filtered to obtain 1-(5-(2-fluorophenyl)-4-methoxy-1-(pyridin-2-ylsulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine as a yellow oil (17 mg, 28%) [M+H].sup.+: 376.

[0247] The compounds listed in Table 1 were synthesized in the same or similar manner to those described above, using appropriate commercially available starting materials and intermediates. The prepared intermediates and Examples were purified using methods well known to those skilled in the art, wherein the methods are not limited to silica gel chromatography, recrystallization, and the like. Further, the final compound obtained from the reaction mixture may be isolated as a neutral, acid or base salt.

TABLE-US-00001 TABLE 1 HPLC NMR Retention LC-MS Intermediate Compound Chemical Time Value (Reaction Example No. Name Shift (Min) [M + H].sup.+ Route) 1 1-(5-(2-Fluorophenyl)-4-methoxy-1- .sup.1H NMR (400 MHz, CDCI.sub.3) 9.067 405 1 ((6-methoxypyridin-3-yl)sulfonyl)- δ 8.12 (d, J = 2.4 Hz, 1H), (Reaction 1H-pyrrol-3-yl)-N-methylmethanamine 7.67-7.65 (m, 2H), 7.47-7.43 (m, 1H), Scheme 1) 7.25 (dt, J = 7.3, 1.7 Hz, 1H), 7.18 (t, J = 7.4 Hz, 1H), 7.05 (t, J = 8.8 Hz, 1H), 6.73 (d, J = 8.8 Hz, 1H), 3.99 (s, 3H), 3.94 (s, 2H), 3.46 (s, 3H), 2.64 (s, 3H). 2 1-(5-(2,4-Difluorophenyl)-4-methoxy-1- .sup.1H NMR (400 MHz, CDCI.sub.3) 9.495 423 3 ((6-methoxypyridin-3-yl)sulfonyl)- δ 8.16 (d, J = 2.4 Hz, 1H), 7.60 (dd, (Reaction 1H-pyrrol-3-yl)-N-methylmethanamine J = 9.0, 2.6 Hz, 1H), 7.46 (s, 1H), Scheme 1) 7.26-7.19 (m, 1H), 6.93 (dt, J = 8.2, 1.7 Hz, 1H), 6.82 (dt, J = 9.0, 2.7 Hz, 1H), 6.73 (d, J = 9.2 Hz, 1H), 4.00 (s, 3H), 3.78 (s, 2H), 3.48 (s, 3H), 2.57 (s, 3H). 3 1-(5-(2,4-Difluorophenyl)-4-methoxy-1- .sup.1H NMR (400 MHz, CDCI.sub.3) 8.593 408 3 ((6-methylpyridin-3-yl)sulfonyl)- δ 8.44 (d, J = 2.0 Hz, 1H), 7.73 (dd, (Reaction 1H-pyrrol-3-yl)-N-methylmethanamine J = 8.4, 2.4 Hz, 1H), 7.60 (s, 1H), 7.22 (d, Scheme 1) J = 8.4 Hz, 1H), 7.15 (q, J = 7.7 Hz, 1H), 6.99 (brs, 1H), 6.89 (dt, J = 8.2, 2.0 Hz, 1H), 6.80 (dt, J = 8.8, 2.4 Hz, 1H), 3.84 (s, 2H), 3.43 (s, 3H), 2.59 (s, 3H), 2.56 (s, 3H). 4 1-(5-(2-Fluorophenyl)-4-methoxy-1- .sup.1H NMR (400 MHz, CDCI.sub.3) 8.453 390 2 ((6-methylpyridin-3-yl)sulfonyl)- δ 8.42 (d, J = 2.4 Hz, 1H), 7.61 (dd, (Reaction 1H-pyrrol-3-yl)-N-methylmethanamine J = 8.4, 2.4 Hz, 1H), 7.34-7.12 (m, 1H), Scheme 2) 7.37 (s, 1H), 7.23-7.13 (m, 3H), 7.04 (t, J = 8.8 Hz, 1H), 3.68 (s, 2H), 3.44 (s, 3H), 2.61 (s, 3H), 2.50 (s, 3H). 5 1-(5-(2,4-Difluorophenyl)-4-methoxy-1- .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 7.92 (t, 8.793 407 3 ((6-methylpyridin-2-yl)sulfonyl)- J = 8.0 Hz, 1H), 7.57 (d, (Reaction 1H-pyrrol-3-yl)-N-methylmethanamine J = 7.6 Hz, 1H), 7.39 (d, Scheme 1) J = 7.6 Hz, 1H), 7.31 (s, 1H), 7.23-7.14 (m, 2H), 7.08-7.05 (m, 1H), 3.54 (s, 2H), 3.43 (s, 3H), 2.48 (s, 3H), 2.32 (s, 3H). 6 1-(5-(2-Fluorophenyl)-4-methoxy-1- .sup.1H NMR {400 MHz, CDCI.sub.3) 8.407 376 2 (pyridin-2-ylsulfonyl)-1H-pyrrol- δ 8.62 (dd, J = 1.6, 5.3 Hz, (Reaction 3-yl)-N-methylmethanamine 1H), 7.77-7.72 (m, 1H), 7.50-7.31 (m, 4H), Scheme 2) 7.21-7.03 (m, 2H), 6.98 (dd, J = 8.8, 8.8 Hz, 1H), 3.70-3.67 (m, 2H), 3.44 (s, 3H), 2.49 (s, 3H).

[0248] In the following Test Examples, experiments were performed using any one or more of the compounds of Examples 1 to 4 according to the present disclosure.

[Test Example 1] Inhibitory Activity on Proton Pump (H.SUP.+./K.SUP.+.-ATPase)

[0249] The proton pump (H+/K+-ATPase) inhibitory activity of the prepared compound was measured as follows. Gastric vesicles isolated from pig stomach were prepared according to the document (see Methods Mol Biol. 2016; 1377:19-27). The protein contents of the gastric vesicles were quantified with a Bicinchoninic Acid (BCA) kit (Sigma Aldrich, BCA1). To each well of a 96-well plate, 70 μl of 50 mM Tris-HEPES buffer (pH 6.5) containing 125 ng of vesicles, DMSO or a substance for each concentration (final DMSO concentration of 1%), 5 mM MgCl.sub.2, and 10 mM KCl was added and pre-incubated at 37° C. for 30 minutes. Then, 10 μl of 2 mM ATP was added to each well, followed by enzymatic reaction at 37° C. for 40 minutes. The reaction was stopped by adding 20 μl of malachite green reagent (Sigma Aldrich, MAK307) and the resulting mixture was allowed to stand at room temperature for 30 minutes. By measuring the absorbance at 620 nm using a Microplate Reader (Biotek, Synergy H4), an amount of inorganic phosphorus released from ATP degradation was measured and the enzyme activity was measured. The absorbance of the enzyme-reacted sample without adding KCl was measured, and the measurement value was subtracted from all the above measurements. Assuming that the group treated with 1% DMSO (DMSO control group) was 100% H.sup.+/K.sup.+-ATPase enzyme activity, and that the group without KCl (KCl control group) was 0% H.sup.+/K.sup.+-ATPase enzyme activity, % inhibition was calculated as in the following Equation 1:

% inhibition=[1−(OD.sub.treatment group−O.sub.DKCl control group)/(OD.sub.DMSO control group−OD.sub.KCl control group))]*100.  [Equation 1]

[0250] IC.sub.50 was obtained by nonlinear regression analysis of GraphPad Prism7 program using % inhibition values for each concentration, and results thereof are shown in Table 2 below.

TABLE-US-00002 TABLE 2 Example IC.sub.50 (μM) 1 +++ 2 +++ 3 +++ 4 +++ (* IC.sub.50 of 0.3 μM or less was indicated by +++)

[0251] As could be seen above, the excellent inhibitory activity of the compound according to the present disclosure on the proton pump (H.sup.+/K.sup.+-ATPase) was confirmed.

[Test Example 2] Inhibitory Activity and Reversibility Evaluation on Proton Pump (H+/K+-ATPase) Depending on pH

[0252] In order to measure the change in proton pump (H+/K+-ATPase) inhibitory activity depending on the pH of the prepared compound, an experiment was performed in the same manner as in Test Example 1 under three conditions of pH 6.5, pH 7.0, and pH 7.5. Examples 1 and 3 were confirmed to have a higher inhibition ability in a weakly acidic condition compared to a neutral condition. It shows that the degree of inhibition ability on the gastric acid pump is better in acidic conditions, and the inhibition ability is restored after the pH in the stomach is restored.

[0253] In addition, in order to confirm the reversibility of the inhibition ability on the proton pump (H.sup.+/K.sup.+-ATPase) of the prepared compound, an experiment was performed by the jump dilution method. 6.25 μg of gastric vesicles isolated from pig stomach and 0.2 μM of each compound were pre-incubated for 120 minutes, and then the enzyme activity before dilution and the enzyme activity after 50-fold dilution were compared for each reaction time, and the reversibility was evaluated. In both Example 1 and Example 3, the inhibition ability of 50% or more was confirmed after 20 minutes of the reaction. On the other hand, when the reaction was performed for 60 minutes after the 50-fold dilution, in both Examples 1 and 3, the enzyme activity was recovered to 90% or more, and reversible results were confirmed.

[0254] Strong gastric acid suppression results in an increase in serum gastrin by a compensatory mechanism, which is highly related to the risk of hypergastrinemia, and the like.

[0255] However, the compounds of Examples 1 and 3 showed inhibitory activity by acting on the proton pump within a short time at low pH, and then showed reversibility of rapidly recovering the enzyme activity.

[0256] This shows a reversible characteristic of restoring acid secretion by easy dissociation from the proton pump, and shows low incidence of hypergastrinemia.

[0257] In other words, it could be expected from the experimental results of the present disclosure that the compounds according to the present disclosure had an excellent effect on inhibiting acid secretion without side effects on hypergastrinemia.

TABLE-US-00003 TABLE 3 pH 6.5 pH 7.0 pH 7.5 Example 1 +++ +++ ++ Example 3 +++ ++ ++ (* IC.sub.50 of 0.1 μM or less was indicated by +++ and IC.sub.50 of greater than 0.1 μM and less than or equal to 1 μM was indicated by ++)

TABLE-US-00004 TABLE 4 % Inhibition Reaction time 20 min 60 min 120 min Before After Before After Before After Dilution Dilution Dilution Dilution Dilution Dilution Dilution Example 1 +++ ++ +++ + +++ + Example 3 +++ ++ +++ + +++ + (* % inhibition of 50% or more was indicated by +++, % inhibition of 20% or more and less than 50% was indicated by ++, and % inhibition of less than 20% was indicated by +)

[Test Example 3] Evaluation on SSTR4 Agonist Effect (Camp Assay)

[0258] The agonism effect on SSTR4 was confirmed by cell-based cAMP functional assay. CHO cells in which human SSTR4 was stably expressed were treated with test substances for each concentration and reacted at 37° C. for 30 minutes, and an amount of the produced cAMP was measured by the HTRF detection method. The % response compared to the reference control agonist (sst-14, 10 nM) was calculated, and the EC.sub.50 was calculated through the concentration-response curve. Results thereof are shown in Table 5 below.

TABLE-US-00005 TABLE 5 Example EC.sub.50 (μM) 1 0.78 3 3.9

[0259] As could be seen above, the compounds according to the present disclosure exhibited excellent effects as SSTR4 agonists.

[Test Example 4] Evaluation on Inhibition Ability of Basal Gastric Acid Secretion in Pylorus-Ligated Rat

[0260] The inhibitory efficacy of the prepared compound on basal gastric acid secretion was measured by employing the Shay's rat model [Shay H, et al., Gastroenterology, 1945, 5, 43˜61].

[0261] Male Sprague Dawley (SD) rats were divided into 8 rats per group, and fasted with only water supply for 24 hours. Then, one hour before pylorus ligation, the control group was orally administered with 0.5% methylcellulose solution, and the other groups were orally administered with the Example compound at a dose of 10 mg/10 mL/kg suspended in 0.5% methylcellulose solution.

[0262] After 5 hours of ligation, the rats were sacrificed under Zoletil and Xylazine anesthesia, and the gastric contents were enucleated by incision of the abdominal cavity. The obtained contents were centrifuged at 3,000 rpm for 10 minutes to separate only the supernatant, and gastric juice was collected. 1 mL of the collected gastric juice was taken into a beaker and the pH was measured using an electrode pH meter. To 1 mL of gastric juice, 0.03 mL each of 0.5% dimethylaminoazobenzene alcohol solution and 1% phenolphthalein alcohol solution was added to make red color, and then 0.1N NaOH solution was added, wherein the volume until the rose tint appeared was determined as the total acidity, and the total acid output was obtained by multiplying the acidity of gastric juice by the amount of gastric juice. The % inhibitory activity of the Example compound was calculated according to the following Equation 2, and results thereof are shown in Table 6 below.

% inhibitory activity of Example compound=[(total gastric acid secretion in Control group−total gastric acid secretion in group treated with Example compound)/total gastric acid secretion in Control group]×100  [Equation 2]

TABLE-US-00006 TABLE 6 pH Inhibitory Activity (%) Vehicle Control 1.74 ± 0.30 Group Example 1 8.25 ± 0.16 +++ Example 2 8.04 ± 0.16 +++ Example 3 8.08 ± 0.24 +++ 90% or more: +++, 80% or more and less than 90%: ++, 70% or more and less than 80%: +

[Test Example 5] Evaluation on Gastric Acid Secretion Inhibition Ability in Lumen-Perfused Rat (LPR)

[0263] The inhibitory efficacy of the prepared compound on histamine-stimulated gastric acid secretion was measured in lumen-perfused rat (LPR) models employing Ghosh & Schild's method [Ghosh M N, et al., Br J Pharmacol Chemother., 1958, 13(1), 54˜61].

[0264] A silicone tube was inserted between the stomach and esophagus of fasted male Sprague Dawley (SD) rats, and physiological saline was allowed to perfuse at the same rate. In addition, the silicone tube was inserted between the pylorus and the duodenum to allow the perfusate that had passed through the stomach to come out. Then, histamine was injected at the same rate through a syringe pump to stabilize the pH in the stomach to about 2.5. After pH stabilization, the control group was administered with only 0.5% methyl cellulose through the jugular vein or duodenum, and the PPI control group was administered with omeprazole, esomeprazole, lansoprazole, rabeprazole, or the like. The other groups were injected with the Example compound by the same route. The perfusate was collected by 7.5 mL aliquots every 15 minutes after drug administration and the pH was measured.

[Test Example 6] Evaluation on Gastric Damage Inhibitory Efficacy in Rat of Indomethacin-Induced Gastric Damage

[0265] The experiment was performed as follows to evaluate the gastric ulcer inhibitory efficacy of the Example compound in rat models of gastric damage induced by indomethacin, a drug in the NSAID classes.

[0266] Among fasted male Sprague Dawley rats, the control group was orally administered with 0.5% methylcellulose solution, and the other groups were orally administered with the Example compound at a dose of 10 mg/10 mL/kg suspended in 0.5% methylcellulose solution.

[0267] In 1 hour after oral administration of the Example compound, indomethacin was orally administered, and after 5 hours, the test animals were sacrificed and the stomach was enucleated. After washing the enucleated surface of the stomach, the greater curvature of the stomach was incised. The incised stomach was spread out and fixed. Then, the ratio of the gastric damaged area was obtained by the total gastric area and the damaged area on the gastric mucosa surface using ImageJ software (NIH, Bethesda), and the % inhibitory activity of the Example compound was calculated according to the following Equation 3. Results thereof are shown in Table 7 below:

% inhibitory activity of Example compound=[(ratio of gastric damaged area in control group−ratio of gastric damaged area in group treated with Example compound)/(ratio of gastric damaged area in control group)]×100  [Equation 3]

TABLE-US-00007 TABLE 7 Inhibitory Activity (%) Dose 10 mg/kg Example 1 +++ Example 3 +++ 97% or more: +++, 90% or more and less than 97%: ++, 80% or more and less than 90%: +

[Test Example 7] Efficacy Evaluation for Ethanol-induced gastric damage and Gastrointestinal Inflammatory Disease

[0268] Alcohol may directly cause damage and bleeding to the gastric mucosal layer, and indirectly promotes the secretion of inflammatory cytokines, lipopolysaccharides, endotoxins, or free radicals through infiltration of macrophages and neutrophils, causing both gastric ulcer and gastrointestinal inflammation. The following experiment was performed to evaluate the gastric ulcer inhibitory efficacy and gastrointestinal anti-inflammatory efficacy of the Example compound in rat models of alcohol-induced gastric damage and gastrointestinal inflammatory disease.

[0269] Among fasted male Sprague Dawley rats, the control group was orally administered with 0.5% methylcellulose solution, and the other groups were orally administered with the Example compound suspended in 0.5% methylcellulose solution.

[0270] In 1 hour after oral administration of the Example compound, the test animals were orally administered with 100% ethanol, anesthetized 1 hour later, and was subjected to laparotomy to collect blood from the posterior vena cava. The blood was allowed to stand at room temperature for about 15 minutes to coagulate, and then centrifuged to separate the serum. After completion of blood collection, the stomach was enucleated. After washing the surface of the enucleated stomach with physiological saline, the greater curvature of the stomach was incised. The incised stomach was placed on a fixture, spread out using forceps, and fixed with a fixing pin. Then, the total gastric area and the damaged area on the gastric mucosa surface were analyzed using ImageJ software (NIH, Bethesda). The gastric tissue was homogenized and centrifuged to obtain gastric tissue protein from the supernatant, and the inflammatory cytokine concentration in the gastric tissue was measured. The inflammatory cytokine concentration in the blood in the isolated serum were measured by enzyme-linked immunosorbent assay (ELISA) technique.

[Test Example 8] Efficacy Evaluation for Acid Reflux-Induced Reflux Esophagitis

[0271] The following experiment was performed to evaluate the esophageal damage inhibitory efficacy of the Example compound in rats with acid reflux-induced reflux esophagitis.

[0272] Among fasted male Sprague Dawley rats, the control group was orally administered with 0.5% methylcellulose solution, and the other groups were orally administered with the Example compound suspended in 0.5% methylcellulose solution.

[0273] In 1 hour after oral administration of the Example compound, the test animals were anesthetized and was subjected to laparotomy. The pylorus of the stomach and the boundary between the proximal stomach and the body were further ligated to allow gastric acid to reflux into the esophagus. After a predetermined period of time, the stomach and esophagus of the test animals were carefully enucleated, the gastric contents were collected and gastric juice was taken, and the pH and amount of gastric juice were measured. The enucleated esophagus was incised in the longitudinal direction and fixed to expose the mucosal area. Esophageal damaged area was analyzed using ImageJ software (NIH, Bethesda).

[Test Example 9] Efficacy Evaluation on Mepirizole-Induced Duodenal Damage

[0274] The experiment was performed as follows to evaluate the duodenal ulcer inhibitory efficacy of the Example compound in rat models of duodenal damage induced by mepirizole, a drug in the NSAID classes.

[0275] Among male Sprague Dawley rats, the control group was orally administered with 0.5% methylcellulose solution, and the other groups were orally administered with the Example compound suspended in 0.5% methylcellulose solution.

[0276] In 1 hour after oral administration of the Example compound, mepirizole was orally administered, and after a predetermined period of time, the test animals were sacrificed and the duodenum was enucleated. After washing the surface of the enucleated duodenum with physiological saline, the damaged area was analyzed using ImageJ software (NIH, Bethesda).

[Test Example 10] Measurement of Change in Gastrin in Blood after Administration of Example Compound

[0277] The experiment was performed as follows to observe the change in gastrin in the blood after administration of the Example compound according to the present disclosure.

[0278] Among fasted male Sprague Dawley rats, the control group was orally administered with 0.5% methylcellulose solution, and the other groups were orally administered with the Example compound suspended in 0.5% methylcellulose solution.

[0279] About 0.5 mL of blood was collected from the jugular vein of the test animals in 5 hours, 8 hours, 12 hours or 24 hours after oral administration of the Example compound. The gastrin concentration in the blood was measured using the ELISA technique in the collected blood.

[Test Example 11] Evaluation on Anti-Inflammatory Efficacy on Indomethacin-Induced Small Intestinal Inflammation

[0280] The following experiment was performed to measure the inflammatory change after administration of the Example compound in rats of small intestinal inflammation induced by indomethacin, a drug in the NSAID classes.

[0281] Among male C.sub.57BL/6 mice or male Sprague Dawley rats, the control group was intraperitoneally administered with 0.5% methylcellulose solution, and the other groups were intraperitoneally administered with the Example compound suspended in 0.5% methylcellulose solution daily for a predetermined period of time. Indomethacin was orally administered on the last day of administration of the Example compound to induce small intestinal inflammation.

[0282] After a predetermined period of time, the test animals were sacrificed and the small intestine was enucleated. After washing the surface of the enucleated small intestine with physiological saline, damage to the small intestine such as bleeding and inflammation, or the like, was analyzed through histological analysis. The enucleated small intestine tissue was homogenized and centrifuged, and then total RNA was obtained from the small intestine tissue in the supernatant, and an amount of inflammatory cytokine mRNA in the small intestine tissue was measured.

[Test Example 12] Gastrointestinal Neuroendocrine Tumor Observation after Long-Term Administration of Example Compound

[0283] The following experiment was performed to observe the degree of gastrointestinal neuroendocrine tumors caused by changes in gastrin secretion after long-term administration of the Example compound.

[0284] Among Sprague Dawley rats, the control group was orally administered with 0.5% methylcellulose solution, and the other groups were orally administered with a high dose of the Example compound suspended in 0.5% methylcellulose solution daily for 2 years. After a predetermined period of time, the test animals were sacrificed, the stomach and duodenum were enucleated and fixed. Then, the degree of hyperplasia of ECL cells and the incidence of neuroendocrine tumors were observed by histopathological analysis, and compared with the control group.

[Test Example 13] Intragastric Distribution Test

[0285] After oral administration of the Example compound in normal rats, the intragastric distribution by time was measured as follows. The prepared compound was dissolved in distilled water containing 0.5% methylcellulose to 0.2 mg/mL, and then orally administered at a dosage of 4 mg/kg. Rats were sacrificed at 1 hour, 6 hours, 12 hours, and 24 hours after administration. Then, blood was exsanguinated through the heart and perfused with physiological saline, the gastric tissue was enucleated, weighed, and stored at −80° C. until the point of analysis. PBS buffer was added so that the ratio of gastric tissue weight to PBS buffer was 1:4, and the compound in gastric tissue was extracted using a homogenizer. The supernatant of the extract was taken and was subjected to protein precipitation using acetonitrile, and then an amount of the Example compound was measured using LC-MS/MS.

[0286] The calculated exposure in the stomach AUC.sub.last, stomach is shown in Table 8 below. Example 1 showed an excellent intragastric distribution, and the concentration in the stomach exceeded ICH of the in vitro H.sup.+/K.sup.+ ATPase inhibition assay at all time points.

TABLE-US-00008 TABLE 8 AUC.sub.last, stomach obtained after a single administration of the compound of Example 1 at a dose of 4 mg/kg AUC.sub.last, stomach (nmol/kg tissue*hr) Compound Example 1 AUC.sub.last, stomach 171, 252

[0287] It was confirmed from the above results that the compounds according to the present disclosure had excellent intragastric distribution effect.

[Test Example 14] Pharmacokinetic Test in Rats and Beagle Dogs

[0288] The Example compound was dissolved in PBS containing 5% DMSO and 20% hydroxypropyl (HP) beta cyclodextrin, and administered intravenously to rats at a dose of 5 mg/kg, and the Example compound was suspended in distilled water containing 0.5% methylcellulose, and orally administered to rats at a dose of 10 mg/kg. The Example compound was dissolved in PBS containing 5% DMSO and 20% hydroxypropyl (HP) beta cyclodextrin, and administered intravenously to beagle dogs at a dose of 5 mg/kg, and the Example compound was suspended in distilled water containing 0.5% methylcellulose, and orally administered to beagle dogs at a dose of 10 mg/kg. Blood samples were collected at scheduled time points after the single intravenous and oral administration of the Example compound to normal rats and beagle dogs. Acetonitrile containing an internal standard material was added to the collected blood sample for protein precipitation. The sample extracted through protein precipitation was centrifuged, and then the supernatant was injected into LC-MS/MS to be subjected to quantitative analysis of the blood concentration of the Example compound. The AUC for each administration route was calculated based on the blood concentration-time profile obtained as a result above, and based on this, the bioavailability (F) upon oral administration was calculated.

[0289] Results thereof are shown in Tables 9 and 10.

TABLE-US-00009 TABLE 9 Pharmacokinetic parameters calculated after single oral administration of Example compounds to rats In vivo Rat PO PK Parameters Oral Dose AUC.sub.inf Compound (mg/kg) (ng*hr/mL) F (%) Example 1 10 252.8 22.0 Example 3 10 275.4 27.3

TABLE-US-00010 TABLE 10 Pharmacokinetic parameters calculated after single administration of Example compound to beagle dogs In vivo Dog PO PK Parameters Oral Dose AUC.sub.inf Compound (mg/kg) (ng*hr/mL) F (%) Example 1 10 10176.3 71.7 Example 3 10 14455.4 83.8

[0290] As could be seen in Tables 9 and 10, the compound according to the present disclosure had very excellent bioavailability (F) upon oral administration to exhibit remarkably excellent effect in view of pharmacokinetics.

[0291] In the present specification, the detailed description of the contents that are able to be sufficiently recognized and inferred by those skilled in the art of the present disclosure has been omitted. In addition to the specific examples described in the present specification, various modifications can be made within the scope that does not change the technical spirit or essential configuration of the present disclosure. Therefore, the present disclosure may be practiced in a manner different from that specifically described and exemplified in the present specification, which can be understood by those skilled in the technical field of the present disclosure.