Analyses were performed in positive ion mode using flow injection mass spectrometry with a mobile phase of 50% aqueous acetonitrile containing 0.1 v/v% formic acid. The immunocytochemical flow cytometric analysis alluded to a cell cycle arrest at G2/M in HeLa cells with cell accumulation in the M phase. An tubulin proved it polymerisation assay that compound 3 significantly increases the maximum rate of microtubule formation. The antimigratory experiment showed that this triazole (3) inhibits the migration and invasion of HeLa cells. Based Carotegrast on these encouraging results, the 1-benzyl-1,2,3-triazol-4-yl moiety was introduced onto C-3-of 13-oestrone bearing an intact ring D16. Our concept was to improve the one-micromolar IC50 value of the best D-secoestrone triazole by synthesising new compounds bearing the same structural element at C-3-substituent with IC50 values in the submicromolar range. These results highlight the importance of 13-oestrone as a scaffold and the 3-substituent (R?=?Positions or H relative to the C-3-function, yielding the two regioisomers in a ratio of Rabbit Polyclonal to RAB33A 11:12?=?2:1. Bromo derivatives (11a,b or 12a,b) were subjected to Pd-catalysed reactions with diethyl phosphite or diphenylphosphine oxide Carotegrast as coupling partners. Microwave-assisted Hirao couplings afforded new 2- or 4-phosphonated 3-tubulin polymerisation assay in two different concentrations (125 and 250?M). The calculated maximum rate of tubulin polymerisation was increased by our test compound Carotegrast which was significant when 12a was added in 250?M concentration to the reaction mixture (Figure 4). Paclitaxel, the positive control agent recommended by the manufacturer, evoked a threefold increase in microtubule formation. Control: untreated samples. The experiment was performed in two parallels and the measurements were repeated twice. Each bar denotes the mean??SEM, regioisomers in a ratio of 8:9?=?2:1 in high yields. Interestingly, regioselectivity of the bromination depends on the nature and size of the C-3-function markedly. The difference in regioisomeric ratios compared to those of 3-substituent in 3-substituent. The transformations of 3-with a Finnigan TSQ-7000 triple quadrupole mass spectrometer (Finnigan-MAT, San Jose, CA) equipped with a Finnigan electrospray ionisation source. Analyses were performed in positive ion mode using flow injection mass spectrometry with a mobile phase of 50% aqueous acetonitrile containing 0.1 v/v% formic acid. The flow rate was 0.3?ml/min. Five l aliquot of the samples were loaded into the flow. The ESI capillary was adjusted to 4.5?n2 and kV was used as a nebuliser gas. 4.1.1. Synthesis of 3-(prop-2-inyloxy)-13-estra-1,3,5(10)-triene (10) 3-Hydroxy-13-estra-1,3,5(10)-trien-17-one (1, 540?mg, 2.0?mmol) was dissolved in acetone (15?ml), propargyl bromide [0 then.34?ml (80?wt.% in toluene), 3.0?mmol], and K2CO3 (1.94?g, 14?mmol) were added. The reaction mixture was stirred at 70?C for 24?h, the Carotegrast solvent was evaporated Carotegrast off, and the residue was purified by flash chromatography with EtOAc/CH2Cl2 = 2/98 as eluent. Compound 7 was obtained as a white solid (610?mg, 98%), mp 133?134?C, Rf = 0.70 (ss B); Anal calcd. for C21H24O2: C, 81.78; H, 7.84. Found: C, 81.93; H, 7.64. 1H NMR: ppm H 1.06 (s, 3H, H-18); 2.49 (s, 1H, CCH); 2.83 (m, 2H, H-6); 4.65 (s, 2H, OCH2); 6.68 (s, 1H, H-4); 6.77 (d, ppm 1.05 (s, 3H, H-18); 2.80 (m, 2H, H-6); 5.14 (s, 2H, OCH2); 5.51 (s, 2H, NCH2); 6.67 (s, 1H, H-4); 6.75 (dd, ppm 1.05 (s, 3H, H-18); 1.31 (s, 3??3H, C(CH3)3); 2.81 (m, 2H, H-6); 5.11 (s, 2H, OCH2); 5.50 (s, 2H, NCH2); 6.69 (s, 1H, H-4); 6.78 (m, 1H, H-2); 7.16?7.20 (overlapping multiplets, 3H, H-1, H-2, H-6); 7.39 (d, 2H, H-3, H-5); 7.55 (s, 1H, C?=?CH). Compound 4a is identical with the compound described in Ref. [16]. 4.1.3. General procedure for the bromination of triazoles 4a and 4b Triazole 4a or 4b (442?mg or 498?mg, 1.00?mmol) was dissolved in dichloromethane (5?ml) and NBS (178?mg, 1.00?mmol) was added. The mixture was stirred at rt for 2.5?h, the solvent was then evaporated off and the crude product was purified by flash chromatography with EtOAc/hexane = 30/70 as eluent. Synthesis of 3-[{1-benzyl-1ppm: 21.0 (CH2), 25.0.