Ite powder (0.040 g, 77 ) followed by reverse phase flash chromatography (NH2 capped SiO2, three g, 100 CH2Cl2) for biological evaluation: TLC Rf = 0.1 (five MeOH/ CH2Cl2); mp 129.3-131.1 ; 1H NMR (500 MHz, CDCl3) 7.59- 7.55 (m, 4H), 7.48 (d, J = 8. Hz, 2H), 7.42 (dd, J = 7.six, 7.6 Hz, 2H), 7.42 (dd, J = 7.six, 7.six Hz, 1H), 7.36-7.30 (m, 1H), 5.13 (s, 2H), 4.87 (s, 2H), 4.07 (q, J = 7.1 Hz, 1H), two.69 (q, J = 7.six Hz, 2H), 1.61 (d, J = 7.1 Hz, 3H), 1.23 (t, J = 7.six Hz, 3H); 13C NMR (125 MHz, CDCl3) 173.7, 164.five, 160.9, 142.six, 140.9, 140.0, 128.9, 127.6, 127.five, 127.four, 127.three, 101.8, 90.8, 75.six, 32.9, 29.9, 24.9, 12.eight; IR (neat cm-1) 3415, 3304, 3162, 2973, 2927, 2871, 1618, 1547, 1436, 1281, 761, 692, 479; HRMS (ESI, M+ + H) m/z 343.1907 (calculated for C22H23N4, 343.1917). HPLC (a) tR = 19.1 min, 98.9 ; (b) tR = 17.three min, 98.5 . 6-Ethyl-5-[3-(6-phenyl-pyridin-3-yl)-but-1-ynyl]-pyrimidine-2,4diamine (46). As outlined by the common Sonogahisra coupling procedure, ethyl-iodopyrimidine (0.071 g, 0.27 mmol), CuI (0.dx.doi.org/10.1021/jm401916j | J. Med. Chem. 2014, 57, 2643-Journal of Medicinal Chemistryg, 0.06 mmol, 21 mol ), Pd(PPh3)2Cl2 (0.019 g, 0.03 mmol, 10 mol ), and alkyne 43 (0.061 g, 0.three mmol) had been reacted in DMF/Et3N (1 mL every single) at 60 for 12 h. Immediately after the mixture was cooled, the dark reddish brown remedy was TGF alpha/TGFA Protein custom synthesis concentrated, plus the product was purified by flash chromatography (SiO2, five g, two MeOH/CHCl3) to afford Annexin V-FITC/PI Apoptosis Detection Kit Publications coupled pyrimidine 46 as a pale white hygroscopic solid (0.070 g, 75 ), followed by reverse phase flash chromatography (NH2 capped SiO2, 3 g, one hundred CH2Cl2, 1 MeOH/CH2Cl2) for biological evaluation: TLC Rf = 0.1 (5 MeOH/CH2Cl2); 1H NMR (500 MHz, CDCl3) 8.72 (d, J = two.1 Hz, 1H), 7.96 (d, J = 7.2 Hz, 2H), 7.81 (dd, J = eight.2, two.three Hz, 1H), 7.70 (d, J = eight.1 Hz, 1H), 7.46 (dd, J = 7.five, 7.five Hz, 1H), 7.46 (dd, J = 7.five, 7.five Hz, 1H), 7.41-7.38 (m, 1H), 5.09 (s, 2H), four.84 (s, 2H), 4.11 (q, J = 7.1 Hz, 1H), 2.68 (q, J = 7.6 Hz, 2H), 1.63 (d, J = 7.1 Hz, 3H), 1.22 (t, J = 7.6 Hz, 3H); 13C NMR (125 MHz, CDCl3) 173.9, 164.four, 160.9, 156.four, 148.6, 139.3, 137.3, 135.three, 129.1, 128.9, 127.1, 120.6, 100.6, 90.four, 76.2, 30.6, 29.9, 24.7, 12.7; IR (neat cm-1) 3469, 3308, 3166, 2972, 2931,1730, 1542, 1435, 1238, 1018, 739, 692; HRMS (ESI, M+ + H) m/z 344.1865 (calculated for C21H21N5, 344.1875). HPLC (a) tR = six.9 min, 99.5 ; (b) tR = 7.1 min, 99.2 . 6-Ethyl-5-[3-(6-p-tolyl-pyridin-3-yl)-but-1-ynyl]-pyrimidine-2,4-diamine (47). As outlined by the basic Sonogahisra coupling process, ethyl-iodopyrimidine (0.059 g, 0.23 mmol), CuI (0.009 g, 0.05 mmol, 21 mol ), Pd(PPh3)2Cl2 (0.016 g, 0.022 mmol, 10 mol ), and alkyne 44 (0.06 g, 0.27 mmol) have been reacted in DMF/Et3N (1 mL each and every) at 60 for 12 h. Immediately after the mixture was cooled, the dark reddish brown answer was concentrated, plus the item was purified by flash chromatography (SiO2, 5g, two MeOH/CHCl3) to afford coupled pyrimidine 47 as a pale white powder (0.063 g, 76 ) followed by reverse phase flash chromatography (NH2 capped SiO2, 3g, one hundred CH2Cl2, 1 MeOH/CH2Cl2) for biological evaluation: TLC Rf = 0.1 (five MeOH/CH2Cl2); mp 144-146.1 ; 1H NMR (500 MHz, CDCl3) eight.74 (d, J = 2.two Hz, 1H), 7.91 (d, J = 8.1 Hz, 2H), 7.82 (dd, J = 8.2, two.3 Hz, 1H), 7.71 (d, J = 8.two Hz, 1H), 7.30 (d, J = 8.6 Hz, 2H), five.25 (s, 2H), five.07 (s, 2H), four.13 (q, J = 7.1 Hz, 1H), 2.72 (q, J = 7.6 Hz, 2H), 2.42 (s, 3H), 1.66 (d, J = 7.1 Hz, 3H), 1.26 (t, J = 7.six Hz, 3H); 13C NMR (125 MHz, CDCl3) 173.9, 164.five, 161.1, 156.four, 148.five, 139.1.