Table 2.

Validated HNF4α ligands with control experiment HNF4α modulatory activity and binding affinity to the recombinant HNF4α ligand-binding domain (LBD). EC₅₀ and IC₅₀ values were calculated from dose–response curves and are the mean ± SD of at least four biologically independent repeats in duplicates. Fold and remaining (rem.) activation (act.) refer to the maximum fold increase or decrease in reporter activity relative to 0.1% DMSO. Binding was determined by isothermal titration calorimetry (ITC). n.d.—not determined

ID	Structure	HNF4α Ligand Type	Validated HNF4α Modulation	HNF4α LBD Binding
4		Agonist	EC50 15 ± 1 µM 5.6 ± 0.3-fold act.	no/weak binding
5		Agonist	EC50 > 100 µM	n.d.
6		Agonist	EC50 5.8 ± 0.6 µM 6.1 ± 0.7-fold act.	no binding
7		Agonist	EC50 31 ± 8 µM 3.8 ± 0.6-fold act.	Kd 7 µM
9		Inverse agonist	IC50 8 ± 2 µM 0.57 ± 0.04 rem. act.	Kd 0.3 µM
10		Inverse agonist	IC50 24 ± 5 µM 0.65 ± 0.05 rem. act.	Kd 1.7 µM