l DMSO gel To 2 mg ml rat liver microsomes containing added a system of NADPH in a final volume of 500 l of 0.05 M Tris-HCl, pH 7.4. The reaction mixtures were incubated for 3 min at 37 and then the reaction initiated Dovitinib by the addition of compounds. Incubation of contr Was carried out in the absence of coxibs. Incubations were performed in a water bath, acid under stirring for 2 hours at 37 and ending at 100 L 15 trichloroacetic. The reaction mixture was centrifuged at 15,000 g for 15 min and the Cured Walls were analyzed by UPLC MS MS, as described below. 2.8.1. UPLC MS MS samples were prepared as described above using a Acquity LC MS MS system consisting TQ from an ACQUITY UPLC pump, autosampler, an ACQUITY UPLC HSS T3 column BEH 45, with a UV detector diode array spectrometer at Acquity Tandem mass spectrometry in serial mode. UPLC was performed at a flow rate of 0.5 ml min performed using the following conditions: L solvent A was 5 mM ammonium acetate and solvent B is acetonitrile L. Gradient program was L Solvent A in 95 min at 100 0.5 A and 5 BL Solvent L Solvent, followed by a linear gradient from 2.5 to 80 min of L Solvent B conducted and maintained for 1 min 80 L solvent B The injection volume for each sample was 5 L. The Waters Acquity TQ tandem mass spectrometer was equipped with a working electrospray ionization probe in both positive and negative ion mode with a capillary voltage of 2.5 kV. Nitrogen was nebulized for both the heart, and not the gas supplied at flow rates at 20 h and 760 L held or used.
The source and desolvation gas temperatures were 140 and 450th Simple ion scan range was 100 to 800 for the mode both positively and negatively. Scan time was 0.2 s with a delay Delay of 0.02 s inter-scan. 2.9 Effect of sulphaphenazole SB-715992 in theaters selenocoxib 2 To investigate the r Cytochrome P450 in the metabolism selenocoxib 2 RAW 264.7 cells were treated with ketoconazole or sulphaphenazole 2.5M for 30 min after the celecoxib or selenocoxib 2 was added 1 M for 12 h. GPX1 expression in these cells was analyzed by Western immunoblot. DMSO is used as a vehicle in these studies. 2.10. Statistical Analysis Data are expressed as mean SEM and compared between treatment groups with Student’s t-test with GraphPad Prism software program. The criterion for statistical significance was P 0.05. Third RESULTS 3.1. Synthesis selenocoxibs as sulfonamide and pyrazole are essential for the activity T of coxibs, we decided to make use of celecoxib as a molecular platform and made Changes only at positions 3 and 5 Celecoxib has been reported with use of the method. Synthesis selenocoxib 2 in Figure 2A. The key intermediate in this synthesis pyrazole methyl ester 2 was. By reaction of 2, 4, diketone, one made with hydrazine hydrochloride in ethanol with a yield of 77 Ethanol is the L Solvent of choice, that only desired isomer 1.5 as described above. Reduction of the ester group in the compound 2 was shown to be the use of LiAlH4 to hydroxymethyl, 3 in quantitative yield. Chlorinated compound was prepared in a four-pot synthesis by reaction of compound 3 with