Identification and characterization of a phenyl-thiazolyl-benzoic acid derivative as a novel RAR/RXR agonist.

OBJECTIVE: To identify an agonist of RXRα and RARα with reduced undesired profiles of all-trans retinoic acid for differentiation-inducing therapy of acute promyelocytic leukemia (APL), such as its susceptibility to P450 enzyme, induction of P450 enzyme, increased sequestration by cellular retinoic acid binding protein and increased expression of P-glycoprotein, a virtual screening was performed. RESULTS AND CONCLUSION: In this study, a phenyl-thiazolyl-benzoic acid derivative (PTB) was identified as a potent agonist of RXRα and RARα. PTB was characterized in nuclear receptor binding, reporter gene, cell differentiation and cell growth assays. PTB bound directly to RXRα and RARα, but not to PPARα, δ(ß) or γ. PTB fully activated reporter genes with enhancer elements for RXRα/RXRα, and partially activated reporter genes with enhancer elements for RARα/RXRα, PPARδ(ß) and PPARγ. Furthermore, PTB induced differentiation and inhibited the growth of human APL cells. Thus, PTB is a novel dual agonist of RXRα and RARα and works as both a differentiation inducer and a proliferation inhibitor to leukemic cells.

Regulation of expression of the mouse adiponectin gene by the C/EBP family via a novel enhancer region.

Adiponectin plays protective roles against the development of insulin resistance and atherosclerosis. To clarify the regulation of adiponectin gene expression, reporter gene assay by using the several truncated mouse adiponectin 5'-flanking regions was performed after the differentiation of 3T3-L1 preadipocytes. The results indicated that a novel mouse adiponectin enhancer exists in the-2228 to -2066 region. Nuclear proteins from the differentiated adipocytes bound to two DNA fragments, namely, -2153 to -2114 and -2093 to -2054. Both fragments had a common motif, CACAATGC, which was similar to the CCAAT/enhancer binding protein (C/EBP) binding motif. A gel mobility shift assay with anti-C/EBPs antibodies showed that C/EBP alpha, beta, and delta bound to this motif. These data provide the first evidence that the transcriptional activity of the mouse adiponectin gene during adipocyte differentiation is enhanced by the motif in a novel adiponectin enhancer region, via the recruitment of the C/EBPs.