The results are shown in Table 2 and graphically represented in Figure 5. Open in a separate window Figure 5. IC50 in M of compounds 3b, 3c, 3e, 3g, 3j, 7b and 8 on MCF7 and HCT116 cell lines. Table 2. Results of log P and cytotoxic screening of compounds 3b, 3c, 3e, 3g, 3j, 7b and 8 on two cell lines. and positions seemed to make extra H-bonding to the enzyme active site which might account for the higher kinase inhibitory activity exerted by compounds 3e and 3g. On the other hand, SAR study of the pyridothienotriazolopyrimidines 6C8 indicated the following remarks: The unsubstituted triazole derivative 6 showed potent pim-1 inhibition with IC50= 1.08 M. Substitution with methyl group in compound 7a reduced the inhibitory activity. All the new compounds were tested for their pim-1 enzyme inhibitory activity and the most active compounds were further tested for their anti-proliferative activity using two different cell lines MCF7 and HCT116. Experimental part General notes Stuart SMP20 apparatus was used to determine the melting points and they were uncorrected. The IR spectra were recorded on Shimadzu IR 435 spectrophotometer (Kyoto, Japan) and the values were represented in cm?1. The 1H NMR and 13C NMR spectra were recorded on Bruker 400 and 100?MHz spectrophotometer, respectively. TMS was used as an internal standard and the chemical shifts were recorded in ppm on level. Both IR and NMR spectra were carried out at Faculty of Pharmacy, Cairo University or college, Cairo, Egypt. The electron impact mass spectra were recorded on Thermo Scientific ISQLT single quadrapole mass spectrometer. Both mass spectra and elemental analyses were carried out at the regional centre for mycology and biotechnology, Al-Azhar University or college, Cairo, Egypt. All reagents and solvents were purified and dried by standard techniques. 3-Amino-5-bromo-4,6-dimethylthieno[2,3-ppm 2.70 (s, 3H, CH3), 2.85 (s, 3H, CH3), 5.08 (s, 2H, OCH2), 5.80C5.83 (dd, 1H, CH-2, ppm 2.69 (s, 3H, CH3), 2.87 (s, 3H, CH3), 5.88 (s, 1H, CH-2), 7.18 (d, 1H, NH), 7.46C7.57 (m, 4H, Ar-H), 8.59 (s, 1H, NH); 13C NMR (100?MHz, DMSO-d6) ppm 19.9, 26.7 (CH3), 65.4 (CH-2), 110.7, 121.4, 121.6, 124.5, 129.1, 131.5, 141.3, 144.2, 144.8, 157.4, 159.5 (Aromatic C), 161.4 (C=O); MS ppm 2.69 (s, 3H, CH3), 2.87 (s, 3H, CH3), 5.90 (s, 1H, CH-2), 7.20C7.53 (m, 4H, Ar-H), 8.30 (s, 1H, NH), 8.60 (s, 1H, NH); Anal. calcd for C17H13BrClN3OS: C, 48.30; H, 3.10; N, 9.94. Found: C, 48.61; H, 3.28; N, 10.11. 8-Bromo-2-(2,4-dihydroxyphenyl)-7,9-dimethyl-2,3-dihydropyrido[3,2:4,5]thieno[3,2-ppm 2.62 (s, 3H, CH3), 2.80 (s, 3H, CH3), 6.33C6.38 (dd, 1H, CH-2, ppm 2.68 (s, 3H, CH3), 2.84 (s, 3H, CH3), 5.69C5.72 (dd, 1H, CH-2, ppm 19.9, 26.7 (CH3), 66.2 (CH-2), 110.5, 114.5, 115.5, 118.1, 121.3, 124.5, 132.3, 144.5, 144.6, 145.3, 145.6, 157.1, 159.4 (Aromatic C), 161.8 (C=O); MS ppm 2.70 (s, 3H, CH3), 2.87 (s, 3H, CH3), 5.85 (s, 1H, CH-2), 7.12C7.57 (m, 4H, Ar-H), 8.32 (s, 1H, NH), 8.55 (s, 1H, NH); MS ppm 2.69 (s, 3H, CH3), 2.84 (s, 3H, CH3), 3.76 (s, 3H, OCH3), 5.75C5.76 (d, 1H, CH-2), 6.73C7.13 (m, 3H, Ar-H), 7.13 (s, 1H, NH), 8.37 (s, 1H, NH), 9.04 (s, 1H, OH); 13C NMR (100?MHz, DMSO-d6) ppm 19.9, 26.7 (CH3), 56.0 (OCH3), 66.7 (CH-2), 111.1, 111.4, 115.3, 119.6, 121.3, 124.6, 131.8, 144.6, 144.7, 147.0, 147.8, 157.1, 159.5 (Aromatic C), 161.8 TAK-063 (C=O); Anal. calcd for C18H16BrN3O3S: C, 49.78; H, 3.71; N, 9.68. Found: C, 50.02; H, 3.89; N, 9.82. 8-Bromo-2-(4-methoxyphenyl)-7,9-dimethyl-2,3-dihydropyrido[3,2:4,5]thieno[3,2-ppm 2.74 (s, 3H, CH3), 2.85 (s, 3H, CH3), 3.73 (s, 3H, OCH3), 5.82 (s, 1H, CH-2), 6.91C7.45 (m, 4H, Ar-H), 8.22 (d, 1H, NH), 8.46 (s, 1H, NH); MS ppm 2.71 (s, 3H, CH3), 2.87 (s, 3H, CH3), 3.70 (br s, 6H, OCH3), 3.77 (s, 3H, OCH3), 5.81 (s, 1H, CH-2), 6.90C7.00 (m, 3H, Ar-H?+?NH), 8.48 (s, 1H, NH); Anal. calcd for C20H20BrN3O4S: C, 50.22; H, 4.21; N, 8.78. Found: C, 50.49; H, 4.37; N, 8.90. 8-Bromo-7,9-dimethyl-2-(thiophen-3-yl)-2,3-dihydropyrido[3,2:4,5]thieno[3,2-ppm 2.69 (s, 3H, CH3), 2.86 (s, 3H, CH3), 5.89C5.91 (dd, 1H, CH-2), 7.06C7.51 (m, 3H, Ar-H), 8.37 (d, 1H, NH), 8.52 (d, 1H, NH); 13C NMR (100?MHz, DMSO-d6) ppm 19.9, 26.8 (CH3), 63.5 (CH-2), 111.1, 121.3, 123.2, 123.4, 124.8, 127.1, 143.5, 144.4, 144.8, 157.2, 159.4 (Aromatic C), 161.5 (C=O); MS ppm 1.37C1.40 (t, 3H, CH3CH2O, ppm 14.5 (CH3CH2), 19.83, 26.9 (ring CH3), 64.0 (CH3CH2), 90.9, 105.0, 114.5, 122.9, 145.9, 152.1, 158.1, 159.0, 159.9 (Aromatic C and CN); Anal. calcd for C13H12BrN3OS: C, 46.16; H, 3.58; N, 12.42. Found: C, 45.93; H, 3.72; N, 12.69. Synthesis of 8-bromo-4-imino-7,9-dimethylpyrido[3,2:4,5]thieno[3,2-d]pyrimidin-3(4H)-amine (5) Compound 4 (0.67?g, 0.002?mol) was mixed with hydrazine hydrate (99%, 6?ml) in absolute ethanol (10?ml). The mixture was heated under reflux for 10?h, allowed to cool and the product was filtered, dried and crystallised from acetic acid. Yield: 90%; mp: >300?C; IR (cm?1): 3367, 3329, 3294 (NH/NH2), 2951, 2920 (CH aliphatic), 1658 (C?=?N); 1H NMR (400?MHz, DMSO-d6) ppm 2.76 (s, 3H, CH3), 3.15 (s, 3H, CH3), 4.95 (s, 2H, NH2), 8.57 (s, 1H,?=CH), 9.18 (s, 1H,?=NH); Anal. calcd for C11H10BrN5S: C, 40.75; H, 3.11; N, 21.60. Found: C, 40.91; H, 3.24; N, 21.93. Synthesis of 8-bromo-7,9-dimethylpyrido[3,2:4,5]thieno[2,3-e][1,2,4]triazolo[1,5-c]pyrimidine (6) A mixture of compound 5 (0.64?g, 0.002?mol) and formic acid (3?ml) was heated under reflux for 5?h. The reaction mixture was allowed to cool and poured onto ice-cold water (20?ml), then the product was filtered, dried and crystallised from acetic acid. Yield: 73%; mp: 277C278?C; IR (cm?1): 2985,.The background absorbance of the multiwell plates was measured at 690?nm and subtracted from the 450?nm measurement. first published work describing the pim-1 inhibitory activity of pyridothienotriazolopyrimidine derivatives (Figure 3). Open in a separate window Figure 3. Designing pyridothienopyrimidinones as pim-1 inhibitors. All the new compounds were tested for their pim-1 enzyme inhibitory activity and the most active compounds were further tested for their anti-proliferative activity using two different cell lines MCF7 and HCT116. Experimental part General notes Stuart SMP20 apparatus was used to determine the melting points and they were uncorrected. The IR spectra were recorded on Shimadzu IR 435 spectrophotometer (Kyoto, Japan) and the values were represented in cm?1. The 1H NMR and 13C NMR spectra were recorded on Bruker 400 and 100?MHz spectrophotometer, respectively. TMS was used as an internal standard and the chemical shifts were recorded in ppm on scale. Both IR and NMR spectra were carried out at Faculty of Pharmacy, Cairo University, Cairo, Egypt. The electron impact mass spectra were recorded on Thermo Scientific ISQLT single quadrapole mass spectrometer. Both mass spectra and elemental analyses were carried out at the regional centre for mycology and biotechnology, Al-Azhar University, Cairo, Egypt. All reagents and solvents were purified and dried by standard techniques. 3-Amino-5-bromo-4,6-dimethylthieno[2,3-ppm 2.70 (s, 3H, CH3), 2.85 (s, 3H, CH3), 5.08 (s, 2H, OCH2), 5.80C5.83 (dd, 1H, CH-2, ppm 2.69 (s, 3H, CH3), 2.87 (s, 3H, CH3), 5.88 (s, 1H, CH-2), 7.18 (d, 1H, NH), 7.46C7.57 (m, 4H, Ar-H), 8.59 (s, 1H, NH); 13C NMR (100?MHz, DMSO-d6) ppm 19.9, 26.7 (CH3), 65.4 (CH-2), 110.7, 121.4, 121.6, 124.5, 129.1, 131.5, 141.3, 144.2, 144.8, 157.4, 159.5 (Aromatic C), 161.4 (C=O); MS ppm 2.69 (s, 3H, CH3), 2.87 (s, 3H, CH3), 5.90 (s, 1H, CH-2), 7.20C7.53 (m, 4H, Ar-H), 8.30 (s, 1H, NH), 8.60 (s, 1H, NH); Anal. calcd for C17H13BrClN3OS: C, 48.30; H, 3.10; N, 9.94. Found: C, 48.61; H, 3.28; N, 10.11. 8-Bromo-2-(2,4-dihydroxyphenyl)-7,9-dimethyl-2,3-dihydropyrido[3,2:4,5]thieno[3,2-ppm 2.62 (s, 3H, CH3), 2.80 (s, 3H, CH3), 6.33C6.38 (dd, 1H, CH-2, ppm 2.68 (s, 3H, CH3), 2.84 (s, 3H, CH3), 5.69C5.72 (dd, 1H, CH-2, ppm 19.9, 26.7 (CH3), 66.2 (CH-2), 110.5, 114.5, 115.5, 118.1, 121.3, 124.5, 132.3, 144.5, 144.6, 145.3, 145.6, 157.1, 159.4 (Aromatic C), 161.8 (C=O); MS ppm 2.70 (s, 3H, CH3), 2.87 (s, 3H, CH3), 5.85 (s, 1H, CH-2), 7.12C7.57 (m, 4H, Ar-H), 8.32 (s, 1H, NH), 8.55 (s, 1H, NH); MS ppm 2.69 (s, 3H, CH3), 2.84 (s, 3H, CH3), 3.76 (s, 3H, OCH3), 5.75C5.76 (d, 1H, CH-2), 6.73C7.13 (m, 3H, Ar-H), 7.13 (s, 1H, NH), 8.37 (s, 1H, NH), 9.04 (s, 1H, OH); 13C NMR (100?MHz, DMSO-d6) ppm 19.9, 26.7 (CH3), 56.0 (OCH3), 66.7 (CH-2), 111.1, 111.4, 115.3, 119.6, 121.3, 124.6, 131.8, 144.6, 144.7, 147.0, Rabbit polyclonal to ZNF345 147.8, 157.1, 159.5 (Aromatic C), 161.8 (C=O); Anal. calcd for C18H16BrN3O3S: C, 49.78; H, 3.71; N, 9.68. Found: C, 50.02; H, 3.89; N, 9.82. 8-Bromo-2-(4-methoxyphenyl)-7,9-dimethyl-2,3-dihydropyrido[3,2:4,5]thieno[3,2-ppm 2.74 (s, 3H, CH3), 2.85 (s, 3H, CH3), 3.73 (s, 3H, OCH3), 5.82 (s, 1H, CH-2), 6.91C7.45 (m, 4H, Ar-H), 8.22 (d, 1H, NH), 8.46 (s, 1H, NH); MS ppm 2.71 (s, 3H, CH3), 2.87 (s, 3H, CH3), 3.70 (br s, 6H, OCH3), 3.77 (s, 3H, OCH3), 5.81 (s, 1H, CH-2), 6.90C7.00 (m, 3H, Ar-H?+?NH), 8.48 (s, 1H, NH); Anal. calcd for C20H20BrN3O4S: C, 50.22; H, 4.21; N, 8.78. Found: C, 50.49; H, 4.37; N, 8.90. 8-Bromo-7,9-dimethyl-2-(thiophen-3-yl)-2,3-dihydropyrido[3,2:4,5]thieno[3,2-ppm 2.69 (s, 3H, CH3), 2.86 (s, 3H, CH3), 5.89C5.91 (dd, 1H, CH-2), 7.06C7.51 (m, 3H, Ar-H), 8.37 (d, 1H, NH), 8.52 (d, 1H, NH); 13C NMR (100?MHz, DMSO-d6) ppm 19.9, 26.8 (CH3), 63.5 (CH-2), 111.1, 121.3, 123.2, 123.4, 124.8, 127.1, 143.5, 144.4, 144.8, 157.2, 159.4 (Aromatic C), 161.5 (C=O); MS ppm 1.37C1.40 (t, 3H, CH3CH2O, ppm 14.5.Besides, a brief SAR study of the substitution on the 2-aryl moiety was done. SMP20 apparatus was used to determine the melting points and they were uncorrected. The IR spectra were recorded on Shimadzu IR 435 spectrophotometer (Kyoto, Japan) and the values were represented in cm?1. The 1H NMR and 13C NMR spectra were recorded on Bruker 400 and 100?MHz spectrophotometer, respectively. TMS was used as an internal standard and the chemical shifts were recorded in ppm on scale. Both IR and NMR spectra were carried out TAK-063 at Faculty of Pharmacy, Cairo University, Cairo, Egypt. The electron impact mass spectra were recorded on Thermo Scientific ISQLT single quadrapole mass spectrometer. Both mass spectra and elemental analyses were carried out at the regional centre for mycology and biotechnology, Al-Azhar University, Cairo, Egypt. All reagents and solvents were purified and dried by standard techniques. 3-Amino-5-bromo-4,6-dimethylthieno[2,3-ppm 2.70 (s, 3H, CH3), 2.85 (s, 3H, CH3), 5.08 (s, 2H, OCH2), 5.80C5.83 (dd, 1H, CH-2, ppm 2.69 (s, 3H, CH3), 2.87 (s, 3H, CH3), 5.88 (s, 1H, CH-2), 7.18 (d, 1H, NH), 7.46C7.57 (m, 4H, Ar-H), 8.59 (s, 1H, NH); 13C NMR (100?MHz, DMSO-d6) ppm 19.9, 26.7 (CH3), 65.4 (CH-2), 110.7, 121.4, 121.6, 124.5, 129.1, 131.5, 141.3, 144.2, 144.8, 157.4, 159.5 (Aromatic C), 161.4 (C=O); MS ppm 2.69 (s, 3H, CH3), 2.87 (s, 3H, CH3), 5.90 (s, 1H, CH-2), 7.20C7.53 (m, 4H, Ar-H), 8.30 (s, 1H, NH), 8.60 (s, 1H, NH); Anal. calcd for C17H13BrClN3OS: C, 48.30; H, 3.10; N, 9.94. Found: C, 48.61; H, 3.28; N, 10.11. 8-Bromo-2-(2,4-dihydroxyphenyl)-7,9-dimethyl-2,3-dihydropyrido[3,2:4,5]thieno[3,2-ppm 2.62 (s, 3H, CH3), 2.80 (s, 3H, CH3), 6.33C6.38 (dd, 1H, CH-2, ppm 2.68 (s, 3H, CH3), 2.84 (s, 3H, CH3), 5.69C5.72 (dd, 1H, CH-2, ppm 19.9, 26.7 (CH3), 66.2 (CH-2), 110.5, 114.5, 115.5, 118.1, 121.3, 124.5, 132.3, 144.5, 144.6, 145.3, 145.6, 157.1, 159.4 (Aromatic C), 161.8 (C=O); MS ppm 2.70 (s, 3H, CH3), 2.87 (s, 3H, CH3), 5.85 (s, 1H, CH-2), 7.12C7.57 (m, 4H, Ar-H), 8.32 (s, 1H, NH), 8.55 (s, 1H, NH); MS ppm 2.69 (s, 3H, CH3), 2.84 (s, 3H, CH3), 3.76 (s, 3H, OCH3), 5.75C5.76 (d, 1H, CH-2), 6.73C7.13 (m, 3H, Ar-H), 7.13 (s, 1H, NH), 8.37 (s, 1H, NH), 9.04 (s, 1H, OH); 13C NMR (100?MHz, DMSO-d6) ppm 19.9, 26.7 (CH3), 56.0 (OCH3), 66.7 (CH-2), 111.1, 111.4, 115.3, 119.6, 121.3, 124.6, 131.8, 144.6, 144.7, 147.0, 147.8, 157.1, 159.5 (Aromatic C), 161.8 (C=O); Anal. calcd for C18H16BrN3O3S: C, 49.78; H, 3.71; N, 9.68. Found: C, 50.02; H, 3.89; N, 9.82. 8-Bromo-2-(4-methoxyphenyl)-7,9-dimethyl-2,3-dihydropyrido[3,2:4,5]thieno[3,2-ppm 2.74 (s, 3H, CH3), 2.85 (s, 3H, CH3), 3.73 (s, 3H, OCH3), 5.82 (s, 1H, CH-2), 6.91C7.45 (m, 4H, Ar-H), 8.22 (d, TAK-063 1H, NH), 8.46 (s, 1H, NH); MS ppm 2.71 (s, 3H, CH3), 2.87 (s, 3H, CH3), 3.70 (br s, 6H, OCH3), 3.77 (s, 3H, OCH3), 5.81 (s, 1H, CH-2), 6.90C7.00 (m, 3H, Ar-H?+?NH), 8.48 (s, 1H, NH); Anal. calcd for C20H20BrN3O4S: C, 50.22; H, 4.21; N, 8.78. Found: C, 50.49; H, 4.37; N, 8.90. 8-Bromo-7,9-dimethyl-2-(thiophen-3-yl)-2,3-dihydropyrido[3,2:4,5]thieno[3,2-ppm 2.69 (s, 3H, CH3), 2.86 (s, 3H, CH3), 5.89C5.91 (dd, 1H, CH-2), 7.06C7.51 (m, 3H, Ar-H), 8.37 (d, 1H, NH), 8.52 (d, 1H, NH); 13C NMR (100?MHz, DMSO-d6) ppm 19.9, 26.8 (CH3), 63.5 (CH-2), 111.1, 121.3, 123.2, 123.4, 124.8, 127.1, 143.5, 144.4, 144.8, 157.2, 159.4 (Aromatic C), 161.5 (C=O); MS ppm 1.37C1.40 (t, 3H, CH3CH2O, ppm 14.5 (CH3CH2), 19.83, 26.9 (ring CH3), 64.0 (CH3CH2), 90.9, 105.0, 114.5,.Whilst, the cytotoxic activity on HCT116 ranged between 0.37 and 4.82?M. their anti-proliferative activity using two different cell lines MCF7 and HCT116. Experimental part General notes Stuart SMP20 apparatus was used to determine the melting points and they were uncorrected. The IR spectra were recorded on Shimadzu IR 435 spectrophotometer (Kyoto, Japan) and the values were represented in cm?1. The 1H NMR and 13C NMR spectra were recorded on Bruker 400 and 100?MHz spectrophotometer, respectively. TMS was used as an internal standard and the chemical shifts were recorded in ppm TAK-063 on scale. Both IR and NMR spectra were carried out at Faculty of Pharmacy, Cairo University, Cairo, Egypt. The electron impact mass spectra were recorded on Thermo Scientific ISQLT single quadrapole mass spectrometer. Both mass spectra and elemental analyses were carried out at the regional centre for mycology and biotechnology, Al-Azhar University, Cairo, Egypt. All reagents and solvents were purified and dried by standard techniques. 3-Amino-5-bromo-4,6-dimethylthieno[2,3-ppm 2.70 (s, 3H, CH3), 2.85 (s, 3H, CH3), 5.08 (s, 2H, OCH2), 5.80C5.83 (dd, 1H, CH-2, ppm 2.69 (s, 3H, CH3), 2.87 (s, 3H, CH3), 5.88 (s, 1H, CH-2), 7.18 (d, 1H, NH), 7.46C7.57 (m, 4H, Ar-H), 8.59 (s, 1H, NH); 13C NMR (100?MHz, DMSO-d6) ppm 19.9, 26.7 (CH3), 65.4 (CH-2), 110.7, 121.4, 121.6, 124.5, 129.1, 131.5, 141.3, 144.2, 144.8, 157.4, 159.5 (Aromatic C), 161.4 (C=O); MS ppm 2.69 (s, 3H, CH3), 2.87 (s, 3H, CH3), 5.90 (s, 1H, CH-2), 7.20C7.53 (m, 4H, Ar-H), 8.30 (s, 1H, NH), 8.60 (s, 1H, NH); Anal. calcd for C17H13BrClN3OS: C, 48.30; H, 3.10; N, 9.94. Found: C, 48.61; H, 3.28; N, 10.11. 8-Bromo-2-(2,4-dihydroxyphenyl)-7,9-dimethyl-2,3-dihydropyrido[3,2:4,5]thieno[3,2-ppm 2.62 (s, 3H, CH3), 2.80 (s, 3H, CH3), 6.33C6.38 (dd, 1H, CH-2, ppm 2.68 (s, 3H, CH3), 2.84 (s, 3H, CH3), 5.69C5.72 (dd, 1H, CH-2, ppm 19.9, 26.7 (CH3), 66.2 (CH-2), 110.5, 114.5, 115.5, 118.1, 121.3, 124.5, 132.3, 144.5, 144.6, 145.3, 145.6, 157.1, 159.4 (Aromatic C), 161.8 (C=O); MS ppm 2.70 (s, 3H, CH3), 2.87 (s, 3H, CH3), 5.85 (s, 1H, CH-2), 7.12C7.57 (m, 4H, Ar-H), 8.32 (s, 1H, NH), 8.55 (s, 1H, NH); MS ppm 2.69 (s, 3H, CH3), 2.84 (s, 3H, CH3), 3.76 (s, 3H, OCH3), 5.75C5.76 (d, 1H, CH-2), 6.73C7.13 (m, 3H, Ar-H), 7.13 (s, 1H, NH), 8.37 (s, 1H, NH), 9.04 (s, 1H, OH); 13C NMR (100?MHz, DMSO-d6) ppm 19.9, 26.7 (CH3), 56.0 (OCH3), 66.7 (CH-2), 111.1, 111.4, 115.3, 119.6, 121.3, 124.6, 131.8, 144.6, 144.7, 147.0, 147.8, 157.1, 159.5 (Aromatic C), 161.8 (C=O); Anal. calcd for C18H16BrN3O3S: C, 49.78; H, 3.71; N, 9.68. Found: C, 50.02; H, 3.89; N, 9.82. 8-Bromo-2-(4-methoxyphenyl)-7,9-dimethyl-2,3-dihydropyrido[3,2:4,5]thieno[3,2-ppm 2.74 (s, 3H, CH3), 2.85 (s, 3H, CH3), 3.73 (s, 3H, OCH3), 5.82 (s, 1H, CH-2), 6.91C7.45 (m, 4H, Ar-H), 8.22 (d, 1H, NH), 8.46 (s, 1H, NH); MS ppm 2.71 (s, 3H, CH3), 2.87 (s, 3H, TAK-063 CH3), 3.70 (br s, 6H, OCH3), 3.77 (s, 3H, OCH3), 5.81 (s, 1H, CH-2), 6.90C7.00 (m, 3H, Ar-H?+?NH), 8.48 (s, 1H, NH); Anal. calcd for C20H20BrN3O4S: C, 50.22; H, 4.21; N, 8.78. Found: C, 50.49; H, 4.37; N, 8.90. 8-Bromo-7,9-dimethyl-2-(thiophen-3-yl)-2,3-dihydropyrido[3,2:4,5]thieno[3,2-ppm 2.69 (s, 3H, CH3), 2.86 (s, 3H, CH3), 5.89C5.91 (dd, 1H, CH-2), 7.06C7.51 (m, 3H, Ar-H), 8.37 (d, 1H, NH), 8.52 (d, 1H, NH); 13C NMR (100?MHz, DMSO-d6) ppm 19.9, 26.8 (CH3), 63.5 (CH-2), 111.1, 121.3, 123.2, 123.4, 124.8, 127.1, 143.5, 144.4, 144.8, 157.2, 159.4 (Aromatic C), 161.5 (C=O); MS ppm 1.37C1.40 (t, 3H, CH3CH2O, ppm 14.5 (CH3CH2), 19.83, 26.9 (ring CH3), 64.0 (CH3CH2), 90.9, 105.0, 114.5, 122.9, 145.9, 152.1, 158.1, 159.0, 159.9 (Aromatic C and CN); Anal. calcd for C13H12BrN3OS: C, 46.16; H, 3.58; N, 12.42. Found: C, 45.93; H, 3.72; N, 12.69. Synthesis of 8-bromo-4-imino-7,9-dimethylpyrido[3,2:4,5]thieno[3,2-d]pyrimidin-3(4H)-amine (5) Compound 4 (0.67?g, 0.002?mol) was mixed with hydrazine hydrate (99%, 6?ml) in absolute ethanol (10?ml). The mixture was heated under reflux for 10?h, allowed to cool and the product was filtered, dried and crystallised from acetic acid. Yield: 90%; mp: >300?C; IR (cm?1): 3367,.The results are shown in Table 2 and graphically represented in Figure 5. Open in a separate window Figure 5. IC50 in M of compounds 3b, 3c, 3e, 3g, 3j, 7b and 8 on MCF7 and HCT116 cell lines. Table 2. Results of log P and cytotoxic screening of compounds 3b, 3c, 3e, 3g, 3j, 7b and 8 on two cell lines. and positions seemed to make extra H-bonding to the enzyme active site which might account for the higher kinase inhibitory activity exerted by compounds 3e and 3g. On the other hand, SAR study of the pyridothienotriazolopyrimidines 6C8 indicated the following remarks: The unsubstituted triazole derivative 6 showed potent pim-1 inhibition with IC50= 1.08 M. Substitution with methyl group in compound 7a reduced the inhibitory activity. independent window Number 3. Designing pyridothienopyrimidinones as pim-1 inhibitors. All the new compounds were tested for his or her pim-1 enzyme inhibitory activity and the most active compounds were further tested for his or her anti-proliferative activity using two different cell lines MCF7 and HCT116. Experimental part General notes Stuart SMP20 apparatus was used to determine the melting points and they were uncorrected. The IR spectra were recorded on Shimadzu IR 435 spectrophotometer (Kyoto, Japan) and the ideals were displayed in cm?1. The 1H NMR and 13C NMR spectra were recorded on Bruker 400 and 100?MHz spectrophotometer, respectively. TMS was used as an internal standard and the chemical shifts were recorded in ppm on level. Both IR and NMR spectra were carried out at Faculty of Pharmacy, Cairo University or college, Cairo, Egypt. The electron effect mass spectra were recorded on Thermo Scientific ISQLT solitary quadrapole mass spectrometer. Both mass spectra and elemental analyses were carried out in the regional centre for mycology and biotechnology, Al-Azhar University or college, Cairo, Egypt. All reagents and solvents were purified and dried by standard techniques. 3-Amino-5-bromo-4,6-dimethylthieno[2,3-ppm 2.70 (s, 3H, CH3), 2.85 (s, 3H, CH3), 5.08 (s, 2H, OCH2), 5.80C5.83 (dd, 1H, CH-2, ppm 2.69 (s, 3H, CH3), 2.87 (s, 3H, CH3), 5.88 (s, 1H, CH-2), 7.18 (d, 1H, NH), 7.46C7.57 (m, 4H, Ar-H), 8.59 (s, 1H, NH); 13C NMR (100?MHz, DMSO-d6) ppm 19.9, 26.7 (CH3), 65.4 (CH-2), 110.7, 121.4, 121.6, 124.5, 129.1, 131.5, 141.3, 144.2, 144.8, 157.4, 159.5 (Aromatic C), 161.4 (C=O); MS ppm 2.69 (s, 3H, CH3), 2.87 (s, 3H, CH3), 5.90 (s, 1H, CH-2), 7.20C7.53 (m, 4H, Ar-H), 8.30 (s, 1H, NH), 8.60 (s, 1H, NH); Anal. calcd for C17H13BrClN3OS: C, 48.30; H, 3.10; N, 9.94. Found out: C, 48.61; H, 3.28; N, 10.11. 8-Bromo-2-(2,4-dihydroxyphenyl)-7,9-dimethyl-2,3-dihydropyrido[3,2:4,5]thieno[3,2-ppm 2.62 (s, 3H, CH3), 2.80 (s, 3H, CH3), 6.33C6.38 (dd, 1H, CH-2, ppm 2.68 (s, 3H, CH3), 2.84 (s, 3H, CH3), 5.69C5.72 (dd, 1H, CH-2, ppm 19.9, 26.7 (CH3), 66.2 (CH-2), 110.5, 114.5, 115.5, 118.1, 121.3, 124.5, 132.3, 144.5, 144.6, 145.3, 145.6, 157.1, 159.4 (Aromatic C), 161.8 (C=O); MS ppm 2.70 (s, 3H, CH3), 2.87 (s, 3H, CH3), 5.85 (s, 1H, CH-2), 7.12C7.57 (m, 4H, Ar-H), 8.32 (s, 1H, NH), 8.55 (s, 1H, NH); MS ppm 2.69 (s, 3H, CH3), 2.84 (s, 3H, CH3), 3.76 (s, 3H, OCH3), 5.75C5.76 (d, 1H, CH-2), 6.73C7.13 (m, 3H, Ar-H), 7.13 (s, 1H, NH), 8.37 (s, 1H, NH), 9.04 (s, 1H, OH); 13C NMR (100?MHz, DMSO-d6) ppm 19.9, 26.7 (CH3), 56.0 (OCH3), 66.7 (CH-2), 111.1, 111.4, 115.3, 119.6, 121.3, 124.6, 131.8, 144.6, 144.7, 147.0, 147.8, 157.1, 159.5 (Aromatic C), 161.8 (C=O); Anal. calcd for C18H16BrN3O3S: C, 49.78; H, 3.71; N, 9.68. Found out: C, 50.02; H, 3.89; N, 9.82. 8-Bromo-2-(4-methoxyphenyl)-7,9-dimethyl-2,3-dihydropyrido[3,2:4,5]thieno[3,2-ppm 2.74 (s, 3H, CH3), 2.85 (s, 3H, CH3), 3.73 (s, 3H, OCH3), 5.82 (s, 1H, CH-2), 6.91C7.45 (m, 4H, Ar-H), 8.22 (d, 1H, NH), 8.46 (s, 1H, NH); MS ppm 2.71 (s, 3H, CH3), 2.87 (s, 3H, CH3), 3.70 (br s, 6H, OCH3), 3.77 (s, 3H, OCH3), 5.81 (s, 1H, CH-2), 6.90C7.00 (m, 3H, Ar-H?+?NH), 8.48 (s, 1H, NH); Anal. calcd for C20H20BrN3O4S: C, 50.22; H, 4.21; N, 8.78. Found out: C, 50.49; H, 4.37; N, 8.90. 8-Bromo-7,9-dimethyl-2-(thiophen-3-yl)-2,3-dihydropyrido[3,2:4,5]thieno[3,2-ppm 2.69 (s, 3H, CH3), 2.86 (s, 3H, CH3), 5.89C5.91 (dd, 1H, CH-2), 7.06C7.51 (m, 3H, Ar-H), 8.37 (d, 1H, NH), 8.52 (d, 1H, NH); 13C NMR (100?MHz, DMSO-d6) ppm 19.9, 26.8 (CH3), 63.5 (CH-2), 111.1, 121.3, 123.2, 123.4, 124.8, 127.1, 143.5, 144.4, 144.8, 157.2, 159.4 (Aromatic C), 161.5 (C=O); MS ppm 1.37C1.40 (t, 3H, CH3CH2O, ppm 14.5 (CH3CH2), 19.83, 26.9 (ring.