D SiO2, 3 g, 100 Dynamin custom synthesis CH2Cl2, 1 MeOH/ CH2Cl2) to afford coupled pyrimidine 32 as a pale white powder (0.065 g, 78 ); TLC Rf = 0.2 (five MeOH/CH2Cl2); mp 130.9-133.1 ; 1H NMR (500 MHz, CDCl3) 7.73-7.70 (m, 2H), 7.69-7.63 (m, 3H), 7.19 (dd, J = 7.eight, 1.7 Hz, 1H), 7.05 (d, J = 1.7 Hz, 1H), 5.24 (s, 2H), 4.98 (s, 2H), 4.45 (q, J = 7.0 Hz, 1H), three.94 (s, 3H), 2.71 (q, J = 7.6 Hz, 2H), 1.55 (d, J = 7.0 Hz, 3H), 1.24 (t, J = 7.six Hz, 3H); 13C NMR (125 MHz, CDCl3) 173.4, 164.5, 160.eight, 156.8, 145.7, 139.three, 132.eight, 132.five, 128.five, 127.9, 119.9, 119.1, 111.1, 109.6, 101.9, 90.8, 74.8, 55.six, 29.8, 26.9, 23.0, 12.7; IR (neat cm-1) 3464, 3428, 3332, 3188, 3029, 2925, 2775, 2546, 1651, 1548, 1445, 1286, 1008, 735, 557; HRMS (DART, M+ + H) m/z 398.1983, (calculated for C24H24N5O, 398.1981). HPLC (a) tR = 19.2 min, 99.six ; (b) tR = 17.five min, 99.five . Carbamic Acid 4-[3-(2,4-Diamino-6-ethyl-pyrimidin-5-yl)-1methyl-prop-2-ynyl]-3-methoxy-biphenyl-4-yl Ester (33). As outlined by the general Sonogahisra coupling procedure, ethyl-iodopyrimidine (0.055 g, 0.21 mmol), CuI (0.008 g, 0.04 mmol, 21 mol ), Pd(PPh3)2Cl2 (0.015 g, 0.021 mmol, ten mol ), and alkyne 23 (0.092 g, 0.31 mmol) had been reacted in DMF/Et3N (1 mL each and every) at 60 for 12 h. Just after the mixture was cooled, the dark reddish brown remedy was concentrated, as well as the solution was EZH1 supplier purified by flash chromatography (SiO2, 5 g, 2 MeOH/CHCl3) to afford coupled pyrimidine 33 as a pale white powder (0.076 g, 84 ) followed by reverse phase flash chromatography (NH2 capped SiO2, three g, 100 CH2Cl2, 1 MeOH/ CH2Cl2) for biological evaluation: TLC Rf = 0.07 (5 MeOH/ CH2Cl2); 1H NMR (500 MHz, MeOD) 7.53 (d, J = 7.eight Hz, 1H), 7.46 (d, J = 8.6 Hz, 2H), 7.13 (dd, J = 7.eight,1.60, 1H), 7.11 (d, J = 1.3 Hz, 1H), six.85 (d, J = 8.6 Hz, 2H), four.41 (q, J = 6.9 Hz, 1H), three.93 (s, 3H), two.67 (q, J = 7.six Hz, 2H), 1.52 (d, J = 7.0 Hz, 3H), 1.22 (t, J = 7.six Hz, 3H); 13C NMR (125 MHz, MeOD) 173.5, 166.1, 162.two, 158.3, 157.9, 142.7, 133.eight, 130.9, 129.1, 128.9, 119.9, 116.7, 110.1, 103.two, 91.4, 74.9, 56.2, 30.four, 27.9, 23.4, 13.three; IR (neat cm-1) 3477, 3386, 3336, 3195, 2970, 2929, 2873, 2361, 2023, 1603, 1437, 1217, 1027, 813. HRMS (ESI, M+ + Na) m/z 455.1947 (calculated for C24H26N5NaO3, 455.1928). HPLC (a) tR = 6.eight min, 98 ; (b) tR = 8.2 min, 98.7 . 4-[3-(2,4-Diamino-6-ethyl-pyrimidin-5-yl)-1-methyl-prop-2ynyl]-3-methoxy-biphenyl-4-carboxylic Acid Methyl Ester (34). In line with the general Sonogahisra coupling procedure, ethyliodopyrimidine (0.061g, 0.23 mmol), CuI (0.009 g, 0.05 mmol, 21 mol ), Pd(PPh3)2Cl2 (0.016 g, 0.023 mmol, 10 mol ), and alkyne 24 (0.100 g, 0.34 mmol) have been reacted in DMF/Et3N (1 mL every single) at 60 for 12 h. Just after the mixture was cooled, the dark reddish brown option was concentrated, plus the item was purified by flash chromatography (SiO2, 5g, two MeOH/CHCl3) to afford coupled pyrimidine 34 as a pale white powder (0.077 g, 77 ) followed by reverse phase flash chromatography (NH2 capped SiO2, three g, 100 CH2Cl2, 1 MeOH/CH2Cl2): TLC Rf = 0.1 (five MeOH/CH2Cl2); mp 168.2-170.eight ; 1H NMR (500 MHz, CDCl3) eight.08 (d, J = eight.55 Hz, 2H), 7.64-7.60 (m, 3H), 7.21 (dd, J = 7.eight, 1.six Hz, 1H), 7.08 (d, J = 1.5 Hz, 1H), five.15 (s, 2H), 4.84 (s, 2H), 4.43 (q, J = 7.0 Hz, 1H), three.93 (s, 3H), three.92 (s, 3H), 2.70 (q, J = 7.six Hz, 2H), 1.54 (d, J = 7.0 Hz, 3H), 1.23 (t, J = 7.six Hz, 3H); 13C NMR (126 MHz, CDCl3) 173.5, 167.2, 164.five, 160.eight, 156.7, 145.7, 140.2, 131.9, 130.3, 129.two, 128.3, 127.2, 120.0, 109.7, 102.1, 90.9, 74.7, 55.eight, 52.4, 29.9, 26.9, 2.