1H-Pyrazole-3-acetamide, 5-[[7-[3-[ethyl(2-hydroxyethyl)amino]propoxy]-4-quinazolinyl]amino]-N-(3-fluorophenyl)-

• for the dose dependent inhibition of aurora kinase B (AURKB)
• as a component to identify that, replication timing regulatory factor 1 (RIF1) and protein phosphatase 1 (PP1) are critical for the regulation of abscission timing in human cells
• as a component to study the relevance of AURKB as a cancer therapeutic target

722544-46-9

110-73-6

722544-51-6

The general procedure for the synthesis of 2-(5-((7-(3-(3-(ethyl(2-hydroxyethyl)amino)propoxy)quinazolin-4-yl)amino)-1H-pyrazol-3-yl)-N-(3-fluorophenyl)acetamide, using the compound (CAS:722544-46-9) and N-ethyl ethanolamine, was carried out as follows: firstly, in an inert atmosphere (e.g., protected by nitrogen), 2-(3-(3-(3-(3-chloropropoxy)quinazolin-4-yl)amino}-1H-pyrazole-5-yl)-N-(3-fluorophenyl)acetamide was synthesized as follows. {[7-(3-chloropropoxy)quinazolin-4-yl]amino}-1H-pyrazol-5-yl)-N-(3-fluorophenyl)acetamide and N-ethyl ethanolamine were dissolved in N,N-dimethylacetamide under an inert atmosphere (e.g., nitrogen protection). Subsequently, the reaction mixture was heated to 90°C under stirring and maintained at this temperature for 12-16 hours (optimally 12 hours). Upon completion of the reaction, the mixture was cooled to about 85°C and water was slowly added to maintain the temperature between 80-85°C. The temperature of the mixture was adjusted to 80°C and a crystal seed with an expected yield of about 1% was added to induce crystallization. Over the next about 20 hours, the mixture was slowly cooled to 20°C to ensure that the product precipitated in the desired crystalline form and that the crystals were of suitable size for filtration. The product was collected by filtration, washed sequentially with a water/N,N-dimethylacetamide mixture and acetonitrile, and subjected to appropriate de-liquefaction to give the product in hydrated form. Subsequently, the filter cake was slurried with warm acetonitrile (about 40°C) for about 2 hours, filtered again, washed with acetonitrile, and finally dried under vacuum or nitrogen flow to give virtually anhydrous 2-{3-[(7-{3-[ethyl(2-hydroxyethyl)amino]propoxy}- quinazolin-4-yl)amino]-1H-pyrazol-5-yl}-N-(3-fluorophenyl)acetamide as an off-white solid in yields of 85-90%. The product was confirmed by 1H-NMR (DMSO-d6) and mass spectrometry (+ve ESI), the characteristic signals and molecular ion peaks were as follows: 1H-NMR (DMSO-d6): δ 10.55 (s, 1H), 9.45 (br s, 1H), 8.98 (s, 1H), 8.8 (d, 1H), 7.63 (m, 1H), 7.47 (pr d 1H), 7.37 (m, 2H), 7.32 (d, 1H), 6.9 (m, 1H), 6.77 (s, 1H), 4.32 (t, 2H), 3.83 (br s, 2H), 3.76 (t, 2H), 3.35 (m, 2H), 3.25 (m, 4H), 2.25 (m, 2H), 1.25 (t, 3H); MS (+ve ve ESI): m/z 508.4 (M + H)+.