title: Novel method for rapid copper chelation assessment confirmed low affinity of D-penicillamine for copper in comparison with trientine and 8-hydroxyquinolines.
abstract: Copper is an essential trace element involved in many physiological processes. Since disorder of copper homeostasis is observed in various pathologies, copper chelators may represent a promising therapeutic tool. This study was aimed at: 1) formation of an in vitro methodology for screening of copper chelators, and 2) detailed analysis of the interaction of copper with clinically used D-penicillamine (D-PEN), triethylenetetramine (trientine), experimentally tested 8-hydroxyquinolines, and the disodium salt of EDTA as a standard chelator. Methodology based on bathocuproinedisulfonic acid disodium salt (BCS), usable at (patho)physiologically relevant pHs (4.5-7.5), enabled assessment of both cuprous and cupric ions chelation and comparison of the relative affinities of the tested compounds for copper. In the case of potent chelators, the stoichiometry could be estimated too. Clioquinol, chloroxine and EDTA formed very stable complexes with Cu(+)/Cu(2+) at all tested pHs, while copper complexes with trientine were stable only under neutral or slightly acidic conditions. Non-substituted 8-hydroxyquinoline was a less efficient copper chelator, but still unequivocally more potent than D-PEN. Both 8-hydroxyquinoline and D-PEN chelation potencies, similarly to that of trientine, were pH-dependent and decreased with pH. Moreover, only D-PEN was able to reduce cupric ions. Conclusively, BCS assay represents a rapid, simple and precise method for copper chelation measurement. In addition, lower binding affinity of D-PEN compared with 8-hydroxyquinolines and trientine was demonstrated.
authors.affiliation: Department of Pharmacology and Toxicology, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Heyrovského 1203, 500 05 Hradec Králové, Czech Republic.
authors.forename: Michal
authors.initials: M
authors.lastname: Říha
authors.forename: Jana
authors.initials: J
authors.lastname: Karlíčková
authors.forename: Tomáš
authors.initials: T
authors.lastname: Filipský
authors.forename: Kateřina
authors.initials: K
authors.lastname: Macáková
authors.forename: Radomír
authors.initials: R
authors.lastname: Hrdina
authors.forename: Přemysl
authors.initials: P
authors.lastname: Mladěnka
mesh_terms: D003300:Copper / Q000737:chemistry*; D015125:Oxyquinoline / Q000737:chemistry*; D010396:Penicillamine / Q000737:chemistry*; D010618:Phenanthrolines / Q000737:chemistry; D014266:Trientine / Q000737:chemistry*
chemical_list: D010618:Phenanthrolines; D015125:Oxyquinoline; D003300:Copper; D010396:Penicillamine; D014266:Trientine