title: Synthesis, structural evaluation, and estrogen receptor interaction of 2,3-diarylpiperazines.
abstract: To develop novel estrogen receptor (ER) ligands, ring-fused derivatives of the hormonally active (1R,2S)/(1S,2R)-1-(2-chloro-4-hydroxyphenyl)-2-(2,6-dichloro-4-hydroxyphenyl)ethylenediamine 4b were synthesized. (2R,3S)/(2S,3R)-2-(2-Chloro-4-hydroxyphenyl)-3-(2,6-dichloro-4-hydroxyphenyl)piperazine 4 induced ligand-dependent gene expression in MCF-7-2a cells, stably transfected with the plasmid ERE(wtc)luc and was therefore used as a lead structure. The influence of the substitution pattern in the aromatic rings (4-OH (1), 2-F,4-OH (2), 2-Cl,4-OH (3), 2,6-Cl2,3-OH (5), and 2,6-Cl2,4-OH (6)) and the effect of N-ethyl chains on the ER binding and activation of gene expression were studied. The synthesis started from the respective methoxy-substituted (1R,2S)/(1S,2R)-configurated 1,2-diarylethylenediamines 6b to 4b, which were reacted with dimethyl oxalate in order to get 5,6-diarylpiperazine-2,3-diones. Reduction with BH3*tetrahydrofuran and ether cleavage with BBr3 yielded the piperazines 1-6. The N-alkylation of the piperazines 1a-3a, which was employed for obtaining compounds 7-11, was succeeded by acetic anhydride followed by reduction and ether cleavage. Nuclear magnetic resonance (NMR) spectroscopical studies revealed a synclinal conformation of the 1,2-diarylethane pharmacophore and a preference of the substituents at the heterocyclic ring for an equatorial position. This spatial structure prevents an interaction with the ER analogously to that of estradiol (E2). Therefore, the piperazines can displace E2 from its binding site only to a very small extent. Only the N-ethyl (8) and N,N'-diethyl (11) derivatives of piperazine 3 showed relative binding affinity values > 0.1% (8, 0.42%, and 11, 0.17%). Nevertheless, ER-mediated gene activation was verified for the piperazines 4 (20%), 6 (73%), 7 (34%), 8 (74%), and 11 (37%) (concentration, 1 microM; E2, 100% activation) on the MCF-7-2a cell line. O-methylation led to completely inactive compounds and showed the necessity of H bridges from the piperazines to the ER for activating gene expression.
authors.affiliation: Institute of Pharmacy, Free University of Berlin, Königin-Luise Strasse 2+4, D-14195 Berlin, Germany. rgust@zedat.fu-berlin.de
authors.forename: Ronald
authors.initials: R
authors.lastname: Gust
authors.forename: Roland
authors.initials: R
authors.lastname: Keilitz
authors.forename: Kathrin
authors.initials: K
authors.lastname: Schmidt
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D002417:Cattle; D003600:Cytosol / Q000378:metabolism; D047628:Estrogen Receptor alpha; D005260:Female; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D066298:In Vitro Techniques; D008024:Ligands; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D008968:Molecular Conformation; D010879:Piperazines / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D011869:Radioligand Assay; D011960:Receptors, Estrogen / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D020845:Selective Estrogen Receptor Modulators / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D013329:Structure-Activity Relationship; D015533:Transcriptional Activation; D014407:Tumor Cells, Cultured; D014599:Uterus / Q000648:ultrastructure
chemical_list: D047628:Estrogen Receptor alpha; D008024:Ligands; D010879:Piperazines; D011960:Receptors, Estrogen; D020845:Selective Estrogen Receptor Modulators; D008156:Luciferases