| Assay Method Information | |
| | In Vitro Phosphodiesterase Type 5 (PDE5) Inhibition Assay |
| Description: | Phosphodiesterase type 5 (PDE5) inhibition assay of compounds of formula I, III, and IV and the reference drug sildenafil were performed according to the manufacturer's instruction. Briefly, Black 96-well non-binding plates with wells were filled with 5 μ/mL of purified PDE5 from BPS Biosciences. The protein was treated with the substance or the vehicle control immediately, and each assay received 50 nM TAMRAcGMP (Molecular Devices). 1.5 hours at 30° C. were spent incubating the plates. The plates were then incubated for an additional 30 minutes at 30° C. with IMAP FP Phosphodiesterase Evaluation Assay (Molecular Devices) binding reagent added to each well. FP was measured using a Biotek Synergy 4 plate reader under the manufacturer's recommendations. |
| Affinity data for this assay | |
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