Discovery and Optimization
of 1,3,4-Trisubstituted-pyrazolone Derivatives as Novel, Potent, and
Nonsteroidal Farnesoid X Receptor (FXR) Selective Antagonists
LBVS of 12480 in-house compounds, followed by HTRF assay,
resulted in one nonsteroidal compound (11) with antagonistic
activity against FXR (69.01 ± 11.75 μM). On the basis of 11, 26 new derivatives (12a–z) were designed and synthesized accordingly. Five derivatives (12f–g, 12p, 12u, and 12y) showed better antagonistic activities against
FXR than compound 11. Remarkably, the most potent derivative, 12u (8.96 ± 3.62 μM), showed antagonistic capability
approximately 10 times and 8-fold higher than that of the control
(GS) and the starting compound 11, respectively. 12u was further confirmed to have high binding affinity with
FXRαLBD, FXR specificity over six other nuclear receptors, and
potent antagonistic activity against FXR in two cell testing platforms. 12u strongly suppressed the regulating effects of CDCA on
FXR target genes. The therapeutic potential of 12u was
identified by lowering the contents of triglyceride and cholesterol
in human hepatoma HepG2 cells and in the cholesterol-fed C57BL/6 mices.