| Assay Method Information | |
| | PTGR2 Inhibiting Activity Assay |
| Description: | Exemplary compounds thus prepared were evaluated for their efficacy in inhibiting PTGR2. In vitro enzyme activity was measured to determine the half maximal inhibitory concentration (IC50) of compounds of this invention as PTGR2 inhibitors. The reduction of 15-keto-PGE2 by PTGR2 requires NADPH. The decrease of NADPH during the reduction reaction indicates inhibition of PTGR2 and thus is used to calculate the IC50 of a PTGR2 inhibitor. Human PTGR2 recombinant protein was mixed with a PTGR2 inhibitor (i.e., a compound of this invention) in a potassium phosphate buffer having a final concentration of 30 mM and a pH value of 7.3. The resultant mixture was pre-incubated at room temperature for 15 minutes. After the incubation, 15-keto-PGE2 was added to a final concentration of 20 μM, together with NADPH (20 μM, final concentration) and a Glo-NADPH reagent (commercially available from Promega Corporation, Madison, Wis.). After incubating at room temperature for 30 minutes, the signal by luminometer was recorded and used to calculate the IC50 value of a PTGR2 inhibitor |
| Affinity data for this assay | |
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