30 fold greater potency compared to the parent molecule cortisol mineralocorticoid receptor mr activation potency was assessed using a hek 293t cell line containing the uas bla reporter uas b a hek 293t cellsensor u00ae this cell line was stably cotransfected with an expression construct containing u03b2 lactamase cdna under control of the gal4 upstream activator sequence uas and another expression construct encoding for the fusion protein gal4 dbd mr lbd results for one experiment performed in duplicate for 9 compounds and the control compound aldosterone in agonist mode are summarized in table 4 all assays were performed as 10 point dose responses using a half log fold dilution series starting with a maximum compound concentration of 100 nm the compounds were incubated for 16 hours the activation of the fusion protein gal4 dbd mr lbd leads to expression of the reporter u03b2 lactamase which is detected by the conversion of a fret substrate in a ratiometric assay format this functional assay allows for measurement of receptor agonism by compounds and can be used to determine compound potency and selectivity assay reproducibility was determined by calculating z u2032 values for untreated versus maximum stimulation the z u2032 value was greater than 0 6 indicating good reproducibility of the assay format several compounds showed dose dependent stimulation of the mr signaling pathway table 4 a 58 year old male visits his ophthalmologist for a routine check up the physician discovers that the patient exhibits an elevated intraocular pressure and is at high risk for future complications the patient is instructed to apply a topical liquid formulation containing one of the compounds in table 1 once daily to each eye the patient returns for.