Synthesis and structural characterization of a cardioactive biotinylated digoxin analogue.

We have biotinylated the terminal glycose of digoxin by reaction of the periodate-oxidized steroid with biotin hydrazide. A biotinylated product (BD-1) was formed which retained significant digoxin receptor (Na+/K+ ATPase) binding activity. Sustained reaction resulted in a second biotinylated product (BD-2) which showed reduced receptor binding activity. The products were characterized by FAB mass spectroscopy and shown to be the mono- and di-biotinylated digoxin conjugates of the oxidized glycose moiety. These analogues may prove useful in determining the subcellular site of digoxin binding.