Cytotoxic activity of diamide-based benzenesulfonamides 5a, 5b and 5h was evaluated following the the protocol of MTT colorimetric assay, as reported previously [35,36]

Cytotoxic activity of diamide-based benzenesulfonamides 5a, 5b and 5h was evaluated following the the protocol of MTT colorimetric assay, as reported previously [35,36]. 3.2.4. and 5g. 2.2. Biological Evaluation 2.2.1. Carbonic Anhydrase InhibitionThe CA inhibitory activity of all the newly prepared diamide-based benzenesulfonamides 5aCh was evaluated towards the cytosolic isoforms hCA I and II, as well as towards the transmembrane tumor-associated isoforms hCA IX and XII using an applied photophysics stopped-flow instrument for assaying the CA-catalyzed CO2 hydration activity [17]. The inhibitory activities were compared to acetazolamide (AAZ), a clinically used standard CA inhibitor. The following SAR could be derived from the results in Table 1: Table 1 Inhibition data of human CA isoforms hCA I, II, IX and XII for diamide-based benzenesulfonamides 5aCh, determined by stopped-flow CO2 hydrase assay, using acetazolamide (AAZ) as a standard drug. as internal standards. The abbreviations used are as follows: s, singlet; d, doublet; m, multiplet. IR spectra were recorded with a Bruker FT-IR spectrophotometer. Reaction courses and product mixtures were routinely monitored by thin layer chromatography (TLC) on silica gel precoated F254 Merck plates. Unless otherwise noted, all solvents and reagents were commercially available and were used without further purification. Azlactones 3aCg were previously reported [24,25]. 3.1.2. General Procedure for Preparation of Target Diamide-Based Benzenesulfonamides 5aChA mixture of = 8.4 Hz, H-3, H-5 of C6H5), 7.49 (t, 1H, = 8.0 Hz, H-4 of C6H5), 7.51 (d, 2H, = 8.0 Hz, H-3, H-5 of 4-Cl-C6H4), 7.62 (d, 2H, = 8.4 Hz, H-2, H-6 of C6H5), 7.74 (d, 2H, = 8.8 Hz, H-2, H-6 of sulfonamide), 7.85 (d, 2H, = 8.8 Hz, H-3, H-5 of sulfonamide), 7.98 (d, 2H, = 8.0 Hz, H-2, H-6 of 4-Cl-C6H4), 10.15 (s, 1H, NH D2O exchangeable), 10.53 (s, 1H, NH D2O exchangeable); 13C NMR (DMSO-= 2.0 Hz, = 8.4 Hz, H-5 of 2,4(Cl)2-C6H3), 7.46 (t, 1H, = 8.0 Hz, H-4 of C6H5), 7.48 (d, 1H, = 8.0 Hz, H-6 of 2,4(Cl)2-C6H3), 7.55 (d, 2H, = 8.0 Hz, H-3, H-5 of C6H5), 7.71 (s, 1H, H-3 of 2,4(Cl)2-C6H3), 7.75 (d, 2H, = 8.8 Hz, H-2, H-6 of sulfonamide), 7.86 (d, 2H, = 8.8 Hz, H-3, H-5 of sulfonamide), 7.91 (d, 2H, = Rucaparib 7.6 Hz, H-2, H-6 of C6H5), 10.14 (s, 1H, NH D2O exchangeable), 10.57 (s, 1H, NH D2O exchangeable); 13C NMR (DMSO-= 8.0 Hz, H-4 of C6H5), 7.45- 7.52 (m, 2H, H-3, H-5 of C6H5), 7.52C7.58 (m, 2H, H-3, H-5 of 4-Br-C6H4), 7.70, 8.28 (d, 2H, H-2, H-6 of C6H5), 7.74 (d, 2H, H-2, H-6 of sulfonamide), 7.85 (d, 2H, H-3, H-5 of sulfonamide), 7.98 (d, 2H, = 8.0 Hz, H-2, H-6 of 4-Br-C6H4), 10.61 (s, 2H, NH D2O exchangeable); 13C NMR (DMSO-= 7.6, H-3, H-5 of C6H5), 7.31, 8.23 (2d, 2H, = 8.4 Hz, H-2, H-6 of C6H5), 7.39C7.51 (m, 4H, Ar-H of 4-CH3-C6H4), 7.49, 7.53 (2t, 1H, = 8.0 Hz, H-4 of C6H5), 7.74 (d, 2H, H-2, H-6 of sulfonamide), 7.85 (d, 2H, H-3, H-5 of sulfonamide), 10.09 (s, 1H, NH D2O exchangeable), 10.45 (s, 1H, NH D2O exchangeable); 13C NMR (DMSO-= 8.8 Hz, H-3, H-5 of C6H5), 7.07, 8.04 (2d, 2H, = 8.4 Hz, H-2, H-6 of C6H5), 7.18, 7.32 (2s, 1H, olefinic), 7.39C7.45 (m, 4H, H-3, H-5 and H-2, H-6 of 4-OCH3-C6H4), 7.47 (t, 1H, = 8.0 Hz, H-4 of C6H5), 7.49 (s, 2H, NH2 D2O exchangeable), 7.54, 7.72 (2d, 2H, = 8.8 Hz, H-2, H-6 of sulfonamide), 7.58, 7.85 (2d, 2H, = 8.8 Hz, H-3, H-5 of sulfonamide), 10.95 (s, 2H, NH D2O exchangeable); Anal. calcd. for C23H21N3O5S (451.50): C, 61.19; H, 4.69; N, 9.31. Found C, 60.88; Rabbit Polyclonal to NUP107 H, 4.65; N, 9.30. N-(1-(2,4-Dimethoxyphenyl)-3-oxo-3-((4-sulfamoylphenyl)amino)prop-1-en-2-yl)benzamide (5f)Yellow powder (yield 85%), m.p. 245C250 C; IR (KBr, cm?1): 3410, 3294 (NH, NH2), 1701, 1639 (2C=O) and 1369, 1161 (SO2); 1H NMR (DMSO-= 2.4 Hz, = 9.2 Hz, H-5, H-6 of (OCH3)2-C6H3), 7.41C7.47 (m, 4H, H-3, H-5 of C6H4 and NH2 D2O exchangeable), 7.49 (t, 1H, = 8.0 Hz, H-4 of C6H5), 7.55 (s, 1H, olefinic), 7.63 (d, 2H, H-2, H-6 of C6H5), 7.72 (d, 2H, = 8.8 Hz, H-2, H-6 of sulfonamide), 7.85 (d, 2H, = 8.8 Hz, H-3, H-5 of sulfonamide), 10.16 (s, 1H, NH D2O exchangeable), 10.65 (s, 1H, NH D2O exchangeable); Anal. calcd. for C24H23N3O6S (481.52): C, 59.87; H, 4.81; N, 8.73. Found C, 60.09; H, 4.83; N, 8.67. N-(1-(3,4-Dimethoxyphenyl)-3-oxo-3-((4-sulfamoylphenyl)amino)prop-1-en-2-yl)benzamide (5g)Yellow powder (yield 90%), m.p. 250C253 C; IR (KBr, cm?1): 3413, 3292 (NH, NH2), 1701, 1639 (2C=O) and 1369, 1161 (SO2); 1H NMR (DMSO-= 8.0 Hz, H-4 of C6H5), 7.57,.CA Inhibitory AssayAn Applied Photophysics stopped-flow instrument was used for assaying the CA catalysed CO2 hydration activity, as reported earlier [17]. derived from the results in Table 1: Table 1 Inhibition data of human CA isoforms hCA I, II, IX and XII for diamide-based benzenesulfonamides 5aCh, determined by stopped-flow CO2 hydrase assay, using acetazolamide (AAZ) as a standard drug. as internal standards. The abbreviations used are as follows: s, singlet; d, doublet; m, multiplet. IR spectra were recorded with a Bruker FT-IR spectrophotometer. Reaction courses and product mixtures were routinely monitored by thin layer chromatography (TLC) on silica gel precoated F254 Merck plates. Unless otherwise noted, all solvents and reagents were commercially available and were used without further purification. Azlactones 3aCg were previously reported [24,25]. 3.1.2. General Procedure for Preparation of Target Diamide-Based Benzenesulfonamides 5aChA mixture of = 8.4 Hz, H-3, H-5 of C6H5), 7.49 (t, 1H, = 8.0 Hz, H-4 of C6H5), 7.51 (d, 2H, = 8.0 Hz, H-3, H-5 of 4-Cl-C6H4), 7.62 (d, 2H, = 8.4 Hz, H-2, H-6 of C6H5), 7.74 (d, 2H, = 8.8 Hz, H-2, H-6 of sulfonamide), 7.85 (d, 2H, = 8.8 Hz, H-3, H-5 of sulfonamide), 7.98 (d, 2H, = 8.0 Hz, H-2, H-6 of 4-Cl-C6H4), 10.15 (s, 1H, NH D2O exchangeable), 10.53 (s, 1H, NH D2O exchangeable); 13C NMR (DMSO-= 2.0 Hz, = 8.4 Hz, H-5 of 2,4(Cl)2-C6H3), 7.46 (t, 1H, = 8.0 Hz, H-4 of C6H5), 7.48 (d, 1H, = 8.0 Hz, H-6 of 2,4(Cl)2-C6H3), 7.55 (d, 2H, = 8.0 Hz, H-3, H-5 of C6H5), 7.71 (s, 1H, H-3 of 2,4(Cl)2-C6H3), 7.75 (d, 2H, = 8.8 Hz, H-2, H-6 of sulfonamide), 7.86 (d, 2H, = 8.8 Hz, H-3, H-5 of sulfonamide), 7.91 (d, 2H, = 7.6 Hz, H-2, H-6 of C6H5), 10.14 (s, 1H, NH D2O exchangeable), 10.57 (s, 1H, NH D2O exchangeable); 13C NMR (DMSO-= 8.0 Hz, H-4 of C6H5), 7.45- 7.52 (m, 2H, H-3, H-5 of C6H5), 7.52C7.58 (m, 2H, H-3, H-5 of 4-Br-C6H4), 7.70, 8.28 (d, 2H, H-2, H-6 of C6H5), 7.74 (d, 2H, H-2, H-6 of sulfonamide), 7.85 (d, 2H, H-3, H-5 of sulfonamide), 7.98 (d, 2H, = 8.0 Hz, H-2, H-6 of 4-Br-C6H4), 10.61 (s, 2H, NH D2O exchangeable); 13C NMR (DMSO-= 7.6, H-3, H-5 of C6H5), 7.31, 8.23 (2d, Rucaparib 2H, = 8.4 Hz, H-2, H-6 of C6H5), 7.39C7.51 (m, 4H, Ar-H of 4-CH3-C6H4), 7.49, 7.53 (2t, 1H, = 8.0 Hz, H-4 of C6H5), 7.74 (d, 2H, H-2, H-6 of sulfonamide), 7.85 (d, 2H, H-3, H-5 of sulfonamide), 10.09 (s, 1H, NH D2O exchangeable), 10.45 (s, 1H, NH D2O exchangeable); 13C NMR (DMSO-= 8.8 Hz, H-3, H-5 of C6H5), 7.07, 8.04 (2d, 2H, = 8.4 Hz, H-2, H-6 of C6H5), 7.18, 7.32 (2s, 1H, olefinic), 7.39C7.45 (m, 4H, H-3, H-5 and H-2, H-6 of 4-OCH3-C6H4), 7.47 (t, 1H, = 8.0 Hz, H-4 of C6H5), 7.49 (s, 2H, NH2 D2O exchangeable), 7.54, 7.72 (2d, 2H, = 8.8 Hz, H-2, H-6 of sulfonamide), 7.58, 7.85 (2d, 2H, = 8.8 Hz, H-3, H-5 of sulfonamide), 10.95 (s, 2H, NH D2O exchangeable); Anal. calcd. for C23H21N3O5S (451.50): C, 61.19; H, 4.69; N, 9.31. Found C, 60.88; H, 4.65; N, 9.30. N-(1-(2,4-Dimethoxyphenyl)-3-oxo-3-((4-sulfamoylphenyl)amino)prop-1-en-2-yl)benzamide (5f)Yellow powder (yield 85%), m.p. 245C250 C; IR (KBr, cm?1): 3410, 3294 (NH, NH2), 1701, 1639 (2C=O) and 1369, Rucaparib 1161 (SO2); 1H NMR (DMSO-= 2.4 Hz, = 9.2 Hz, H-5, H-6 of.