title: Influence of an aziridine precursor on the in vitro binding parameters of rat and ovine corticosteroid-binding globulin (CBG).
abstract: Aziridines are highly reactive alkylating compounds used in cancer treatment. Salsola tuberculatiformis Botsch., which causes prolonged gestation in sheep and contraception in rats, contains a very labile hydroxy-phenylaziridine or its precursor. A less labile analogue, 2-(4-acetoxyphenyl)-2-chloro-N-methyl-ethylammonium chloride (Compound I), was synthesized and has been shown to be contraceptive in rats and to be stabilized by corticosteroid-binding globulin (CBG). The current study compared the binding parameters of rat and ovine CBG and evaluated the effect of the aziridine precursor, Compound I, on these parameters. Kd and Bmax values of 0.646 and 578 nM for corticosterone binding to rat CBG and 0.577 and 19.8 nM for cortisol binding to sheep CBG, respectively, were measured. In competitive binding studies with rat plasma, Ki values of 3.48 nM, 0.856 nM, 22.2 nM, 722 microM, and > 1,000,000 microM for cortisol, corticosterone, progesterone, Compound I, and synephrine (Compound II), respectively, were found, while in sheep plasma the values were 0.409 nM, 1.78 nM, 5.28 nM, 594 microM, and > 1,000,000 microM, respectively. Concentrations of Compound I equivalent to an effective pharmacological dose resulted in a significant (P < 0.01) decrease in CBG bound corticosterone and a significant (P < 0.01) increase in free corticosterone in rat plasma. In sheep, a similar effect was observed with cortisol. Progesterone binding, however, did not appear to be affected significantly by Compound I in either rat or sheep plasma. Compound I was found to be a competitive inhibitor of glucocorticoid binding to CBG. These results suggest that binding of Compound I to CBG with concomitant displacement of endogenous glucocorticoids, but not progesterone, may be part of the mechanism of action of these phenylaziridine compounds.
authors.affiliation: Department of Biochemistry, University of Stellenbosch, Republic of South Africa. al@maties.sun.ac.za
authors.forename: A
authors.initials: A
authors.lastname: Louw
authors.forename: P
authors.initials: P
authors.lastname: Swart
authors.forename: F
authors.initials: F
authors.lastname: Allie
mesh_terms: D000085:Acetates / Q000494:pharmacology*; D000818:Animals; D001388:Aziridines / Q000494:pharmacology*; D001667:Binding, Competitive; D003276:Contraceptives, Oral / Q000494:pharmacology; D005021:Ethylamines / Q000494:pharmacology*; D005260:Female; D005938:Glucocorticoids / Q000097:blood / Q000378:metabolism; D008024:Ligands; D011374:Progesterone / Q000097:blood / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012756:Sheep; D014018:Tissue Distribution / Q000187:drug effects; D014156:Transcortin / Q000187:drug effects / Q000378:metabolism*; D014439:Tyramine / Q000031:analogs & derivatives
chemical_list: C104564:2-(4-acetoxyphenyl)-2-chloro-N-methylethylamine; D000085:Acetates; D001388:Aziridines; D003276:Contraceptives, Oral; D005021:Ethylamines; D005938:Glucocorticoids; D008024:Ligands; D011374:Progesterone; D014156:Transcortin; D014439:Tyramine