Cellular interactions of auranofin and a related gold complex with RAW 264.7 macrophages

Auranofin (AF) is an orally active chrysotherapeutic agent whose precise mechanism of action with its putative target cell, the macrophage, is not known. In a previous paper, we described a sequential thiol exchange mechanism that explained auranofin's molecular mechanism of interaction with RAW 264.7 cells. To further understand the mode of action of AF and to test the validity of the thiol exchange model, we have continued to study the interactions with macrophages of AF and a related gold complex, triethylphosphine gold chloride (TEPG). Evaluation of the effects of AF and TEPG on RAW 264.7 cells demonstrated that: more gold from TEPG than AF associated with cells over time and with a variety of concentrations; and cellular association of AF and TEPG was temperature dependent. The energy of activation for cell association, the rate-limiting step in the thiol exchange process, was lower for TEPG than AF; cellular association and uptake of both compounds did not require metabolic energy; and efflux of both AF and TEPG was time, temperature, and thiol dependent. Based on these and previous data, we conclude that sequential thiol exchange may be a generic phenomenon for cellular uptake and distribution of thiol reactive gold compounds and that the rate-limiting step is the exchange of either tetraacetylthioglucose (TATG) or chloride for a membrane-localized sulfhydryl group.