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note
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50) determined by plotting fractional inhibition against log compound concentration. Data were fitted to the four parameter logistic equation. Murine 11β-HSD1 enzyme inhibition was assessed in CHO cells stably transfected with the full length murine hsd11b1 gene. Enzyme inhibition was determined as described for human 11β-HSD1 following a 4-h incubation of cells, compound and substrate.
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77953284834
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note
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Male C57BL/6 mice (25-30 g in weight) were group housed and allowed free access to food and water. Compounds was dissolved in 5% DMSO, 3% ethanol, 4 mM cyclodextrin. Animals (n = 3 per group) were dosed intraperitoneally or orally with vehicle or compound at 12-hourly intervals. At 2 h following the third dose mice were killed by cervical dislocation. Liver, adipose and brain samples were removed and frozen until analysis was performed. Inhibition of 11β-HSD1 in each tissue was determined by incubating homogenates with 20 nM tritiated cortisone, 2 mM NADPH and 0.2% glucose in Krebs buffer, pH 7.4. Cortisone and cortisol levels were measured by HPLC and the percentage inhibition determined relative to vehicle treated tissue.
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84972986579
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