Activators of P-glycoprotein: Structure-activity relationships and investigation of their mode of action.

P-glycoprotein (P-gp), a 170 kDa plasma membrane protein, is one of the most relevant ABC transporters involved in the development of multidrug resistance (MDR). Understanding its mechanism of transport as well as its interactions with various substrates are basic requirements for the development of adequate therapeutic approaches to overcome this kind of resistance against a broad spectrum of structurally unrelated cytostatic drugs. P-gp modulators (activators) that exert various effects on the intracellular accumulation of distinct P-gp substrates are useful tools for investigating the interactions between multiple drug binding sites of this transport protein. In this study, a series of 27 different imidazobenzothiazoles and imidazobenzimidazoles structurally related to the known P-gp activators QB102 and QB11 was designed, and their modulating properties were investigated. Most of them were able to stimulate P-gp-mediated efflux of daunorubicin and rhodamine 123 in a concentration-dependent manner, but some compounds also displayed weak inhibitory effects. Additionally, P-gp-mediated efflux of vinblastine and colchicine was inhibited by several compounds. Therefore, we concluded that the novel compounds bind to the H site of P-gp and activate the efflux of specific substrates of the R site in a positive cooperative manner, whereas binding of H-type substrates is inhibited competitively. This hypothesis is confirmed by the observation that the modulators do not influence hydrolysis of ATP or its affinity toward P-gp.