Discovery and Optimization of 1,3,4-Trisubstituted-pyrazolone Derivatives as Novel, Potent, and Nonsteroidal Farnesoid X Receptor (FXR) Selective Antagonists

Abstract

LBVS of 12480 in-house compounds, followed by HTRF assay, resulted in one nonsteroidal compound (<b>11</b>) with antagonistic activity against FXR (69.01 ± 11.75 μM). On the basis of <b>11</b>, 26 new derivatives (<b>12a</b>–<b>z</b>) were designed and synthesized accordingly. Five derivatives (<b>12f</b>–<b>g</b>, <b>12p</b>, <b>12u</b>, and <b>12y</b>) showed better antagonistic activities against FXR than compound <b>11</b>. Remarkably, the most potent derivative, <b>12u</b> (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 <b>11</b>, respectively. <b>12u</b> 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. <b>12u</b> strongly suppressed the regulating effects of CDCA on FXR target genes. The therapeutic potential of <b>12u</b> was identified by lowering the contents of triglyceride and cholesterol in human hepatoma HepG2 cells and in the cholesterol-fed C57BL/6 mices