Anal. Harom), 3.71 (s, 3H, CH3), 2.59 (s, 3H, CH3). MS (ESI): 398 (M + H+). Anal. (C13H12IN5S) C, H, N. 8-Acetyl-4-(314 (M + H+). Anal. (C15H15N5OS) C, H, N. 4-(= 6.8 Hz, CH), 2.55 (s, 3H, CH3), 1.27 (d, 6H, = 6.8 Hz, 2 CH3). MS (ESI): 314 (M + H+). Anal. (C16H19N5S) C, H, N. 4-(= 6.0 Hz, NH), 4.78 (d, 2H, = 6.0 Hz, CH2), 3.15 (hept, 1H, = 7.0 Hz, CH), 2.57 (s, 3H, CH3), 1.32 (d, 6H, = 7.0 Hz, 2 CH3). MS (ESI): 314 (M + H+). Anal. (C16H19N5S) C, H, N. 4-(3-Chloroanilino)-8-(1-methylethyl)-2-methylsulfanylpyrazolo[1,5-= 7.5 Hz, Harom), 7.32 (t, 1H, = 8.1 Hz, Harom), 7.16 (d, 1H, = 7.4 Hz, Rabbit polyclonal to DGCR8 Harom), 3.19 (hept, 1H, = 7.0 Hz, CH), 2.61 (s, 3H, CH3), 1.35 (d, 6H, = 7.0 Hz, 2 CH3). MS (ESI): 334 (M + H+). Anal. (C15H16ClN5S) C, H, N. 4-(= 5.9 Hz, NH), 7.30-7.26 (m, 5H, Harom), 4.82 (d, 2H, = 5.9 Hz, CH2), 3.29 (s, 3H, CH3), 3.19 (hept, 1H, = 7.0 Hz, CH), 1.28 (d, 6H, = 7.0 Hz, 2 CH3). MS (ESI): 346 (M + H+). Anal. (C16H19N5O2S) C, H, N. 4-(3-Chloroanilino)-8-(1-methylethyl)-2-methylsulfonylpyrazolo[1,5-= 8.3 Hz, Harom), 7.18 (d, 1H, = 8.3 Hz, Harom), 3.38 (s, 3H, CH3), 3.29 (hept, 1H, = 7.0 Hz, CH), 1.37 (d, 6H, = 7.0 Hz, 2 CH3). MS (ESI): 366 (M + H+). Anal. (C15H16ClN5O2S) C, H, N. 2-(((= 0.50, CH2Cl2). IR (film): Protosappanin B 3275, 2960, 2875, 1650, 1600, 1560, 1435, 765, 695 cm?1. 1H NMR (300 MHz, CDCl3): 7.56 (s, 1H, H7), 7.33-7.24 (m, 5H, Harom), 7.10 (s, 1H, NH), 5.23 (s, 1H, OH), 4.73-4.65 (m, 2H, CH2), 4.00-3.88 (m, 1H, CH), 3.83 (d, 1H, = 10.8 Hz, CH2O), 3.66 (dd, 1H, = 7.3, 10.8 Hz, CH2O), 3.02 (hept, 1H, = 6.6 Hz, CH), 1.70-1.52 (m, 2H, CH2), 1.28 (d, 6H, = 6.6 Hz, 2 CH3), 1.03 (t, 3H, = 7.4 Hz, CH3). MS (EI+VE): 354 (M+). Anal. (C19H26N6O) C, H, N. 2-(((= 0.50, CH2Cl2). Anal. (C19H26N6O) C, H, N. 2-((1(c = 0.27, CH2Cl2). IR (KBr): 3360, 3310, 2955, 1635, 1615, 1575, 1420, 1070, 760, 680 cm?1. 1H NMR (300 MHz, DMSO-+ D2O 75C): 8.05 (s, 1H, Harom), 7.88 (d, 1H, = 8.0 Hz, Harom), Protosappanin B 7.77 (s, 1H, H7), 7.38 (t, 1H, = 8.0 Hz, Harom), 7.16 (d, 1H, = 8.0 Hz, Harom), 3.83 (q, 1H, = 5.9 Hz, CH), 3.60-3.50 (m, 2H, CH2O), 2.96 (hept, 1H, = 7.0 Hz, CH), 1.92-2.02 (m, 1H, CH), 1.27 (d, 6H, = 7.0 Hz, 2 CH3), 0.94 (d, 3H, = 6.8 Hz, CH3), 0.93 (d, 3H, = 6.8 Hz, 3 CH3). MS (ESI): 389 (M + H+). Anal. (C19H25ClN6O) C, H, N. 7-(1-Methylethyl)-2,4-= 7.0 Hz, CH), 2.66 (s, 3H, CH3), 2.60 (s, 3H, CH3), 1.39 (d, 6H, = 7.0 Hz, 2 CH3). MS (ESI): 255 (M + H+). Anal. (C10H14N4S2) C, H, N. 4-Benzylamino-7-(1-methylethyl)-2-(methylsulfanyl)imidazo[2,1-= 5.6 Hz, CH2), 3.35 (hept, 1H, = 7.0 Hz, CH), 2.55 (s, 3H, CH3), 1.37 (d, 6H, = 7.0 Hz, 2 CH3). MS (ESI): 314 (M + H+). Anal. (C16H19N5S) C, H, N. 4-Benzylamino-7-(1-methylethyl)-2-(methylsulfonyl)imidazo[2,1-= 5.8 Hz, CH2), 3.43 (hept, 1H, = 7.0 Hz, CH), 3.32 (s, 3H, CH3), 1.39 (d, 6H, = 7.0 Hz, 2 CH3). MS (ESI): 346 (M + H+). Anal. (C16H19N5O2S2) C, H, N. 2-(((= 0.50, CHCl3). IR (KBr): 3330, 3220, 3100, 1615, 1570, 1540, 1450, 1370, 1055, 755, 720 cm?1. 1H NMR (300 MHz, CDCl3): 7.34-7.27 (m, 5H, Harom), 7.08 (s, 1H, H6), 6.89 (broad s, 1H, NH), 4.73 (d, 2H, = 5.5 Hz, CH2), 4.57 (d, 1H, = 6.4 Hz, OH), 3.89-3.78 (m, 2H, CH2O + CH), 3.70-3.60 (m, 1H, CH2O), 3.37 (large s, 1H, NH), 3.24 (hept, 1H, = 7.0 Hz, CH), 1.73-1.51 (m, 2H, CH2), 1.35 (d, 3H, = Protosappanin B 7.0 Hz, CH3), 1.34 (d, 3H, = 7.0 Hz, CH3), 1.02 (t, 1H, = 7.4 Hz, CH3). MS (EI + VE): Protosappanin B 354 (M+). Anal. (C19H26N6O) C, H, N. Biology Protein Kinase Assays CDK1/cyclin B (native affinity purified from starfish oocytes), CDK2/cyclin A, CDK7/cyclin H and CDK9/cyclin T (recombinant, indicated in baculovirus-infected insect cells) and CDK5/p25 (recombinant, indicated in E. coli) were assayed in the presence of 15 M ATP as previously explained.16 IC50 values were identified from dose-response curves and are indicated in M. Cell Cultures and Reagents A549, Personal computer3, Protosappanin B Hela and 293T cells were cultured in total medium (DMEM without phenol reddish, supplemented with 10% heat-inactivated fetal calf serum, 2 mM Glutamax-I, 100 IU/mL penicillin, 100 g/mL streptomycin). Raji and U937 cells were cultured in total medium (RPMI 1640 without.