Inhibitory efficacy of bufadienolides on Na⁺,K⁺-pump activity versus cell proliferation

Bufadienolides are cytotoxic drugs that may form the basis for anticancer agents. Due to structural and functional similarity to cardiotonic glycosides, application is restricted. We, therefore, investigated correlation of their putative anticancer effects with inhibition of Na⁺,K⁺pumps. The natural bufalin and three derivatives were tested. The anticancer effects of the drugs were checked by observing their inhibitory effects on proliferation of rat liver cancer cells using MTT assay. Inhibition of Na⁺,K⁺-pump was determined by measuring pump-mediated current of rat α1/β1 and α2/β1 Na⁺,K⁺pumps expressed in Xenopus oocytes. All tested bufadienolides inhibited cell proliferation and Na⁺,K⁺pump activity. An activity coefficient A=100xIC^{Na,K pump}/IC₅₀^{proliferation} was used to describe drug effectivity as anticancer drug. Natural bufalin exhibited lowest effectivity on cell proliferation, and also the A value for rat α1 isoform was the lowest (0.08), the α2 isoform was much less sensitive (A=1.00). The highest A values were obtained for the BF238 derivative with A=0.88 and 2.64 for the α1 and α2 isoforms, respectively. Therefore, we suggest that search for bufalin derivatives with high anticancer effect and low affinity for both Na⁺,K⁺pump isoforms may be a promising strategy for development of anticancer drugs.