反应详情

N-ethyl-N-isopropylpropan-2-amine (0.043 mL, 0.25 mmol) was added to stirring solution of 3-(2-(4-fluorophenyl)-3-(methylcarbamoyl)-6-((2,2,2-trifluoroethyl)amino)furo[2,3-b]pyridin-5-yl)benzoic acid (15 mg, 0.031 mmol), 2-(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)-1,1,3,3-tetramethylisouronium hexafluorophosphate(V) (18 mg, 0.046 mmol) and 1,2,4-Oxadiazole-3-methanamine, α,α,5-trimethyl hydrochloride (5.5 mg, 0.031 mmol) in DMF (1 mL) at rt. The mixture was allowed to stir at rt. for 1 h. LCMS shows complete reaction. The solution was diluted with MeOH, filtered and purified via preparative LC/MS with the following conditions: Column:Waters XBridge C18, 19×100 mm, 5-μm particles; Guard Column:Waters XBridge C18, 19×10 mm, 5-μm particles; Mobile Phase A: water; Mobile Phase B: acetonitrile; Buffer: 20-mM ammonium acetate; Gradient: 20-95% B over 10.9 minutes, then a 4.0 minute hold at 95% B; Flow: 25 mL/min. Fractions containing the desired product were combined and dried via centrifugal evaporation. The yield of the product was 2.3 mg, and its estimated purity by LCMS analysis was 100%. Two analytical LC/MS injections were used to determine the final purity. Injection 1 conditions: Column:Waters BEH C18, 2.0×50 mm, 1.7-μm particles; Mobile Phase A: 5:95 acetonitrile:water with 10 mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile:water with 10 mM ammonium acetate; Temperature: 40° C.; Gradient: 0.5 min hold at 0% B, 0-100% B over 4 minutes, then a 0.5-minute hold at 100% B; Flow: 1 mL/min. Injection 2 conditions: Column:Waters BEH C18, 2.0×50 mm, 1.7-μm particles; Mobile Phase A: 5:95 methanol:water with 10 mM ammonium acetate; Mobile Phase B: 95:5 methanol:water with 10 mM ammonium acetate; Temperature: 40° C.; Gradient: 0.5 min hold at 0% B, 0-100% B over 4 minutes, then a 0.5-minute hold at 100% B; Flow: 0.5 mL/min. Proton NMR was acquired in deuterated DMSO. LC-MS retention time: 1.81 min; m/z (MH+): 611. LC data was recorded on a Shimadzu LC-10AS liquid chromatograph equipped with a Phenomenex-Luna 3u C18 2.0×30 mm column using a SPD-10AV UV-Vis detector at a detector wave length of 220 nM. The elution conditions employed a flow rate of 1 mL/min, a gradient of 100% solvent A/0% solvent B to 0% solvent A/100% solvent B, a gradient time of 2 min, a hold time of 1 min, and an analysis time of 3 min where solvent A was 10% acetonitrile/90% H2O/0.1% trifluoroacetic acid and solvent B was 10% H2O/90% acetonitrile/0.1% trifluoroacetic acid. MS data was determined using a Micromass Platform for LC in electrospray mode. 1H NMR (500 MHz, DMSO-d6) δ 8.64 (br s, 1H), 8.38 (d, J=4.3 Hz, 1H), 8.00-7.84 (m, 4H), 7.68 (s, 1H), 7.63-7.55 (m, 2H), 7.34 (t, J=8.7 Hz, 2H), 6.63 (br s, 1H), 4.18 (m, 2H), 3.32 (m, 3H), 2.79 (d, J=4.3 Hz, 3H), 2.53 (s, 3H), 1.67 (s, 6H).

1. customreaction
2. filtrationfiltered
3. custompurified via preparative LC/MS with the following conditions
4. waitGradient: 20-95% B over 10.9 minutes
5. customa 4.0 minute hold
6. additionFractions containing the desired product
7. customdried via centrifugal evaporation
8. custom40° C.
9. waitGradient: 0.5 min hold at 0% B, 0-100% B over 4 minutes
10. customa 0.5-minute hold
11. custom40° C.
12. waitGradient: 0.5 min hold at 0% B, 0-100% B over 4 minutes
13. customa 0.5-minute hold
14. customequipped with a Phenomenex-Luna 3u C18 2.0×30 mm column
15. washThe elution conditions
16. customa gradient time of 2 min
17. customa hold time of 1 min
18. customan analysis time of 3 min where solvent A was 10% acetonitrile/90% H2O/0.1% trifluoroacetic acid and solvent B