Mass spectrometric behavior of cardiac steroid aglycones of the cardenolide type.
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A simple method is presented for the separation of cardiac glycosides from cardiac aglycones using highly cross-linked lipophilic dextran gel chromatography with 95% ethanol as the solvent.
A series of compounds related to 3-deoxydigitoxigenin was prepared and assayed for inhibition of myocardial Na+,K+- adenosine triphosphatase. Although the relatively high activity of 3-deoxydigitoxin was confirmed, the corresponding 3beta,4beta-epoxide and a mixture of 2,3-olefins and 3,4-olefins were less active. 3-Deoxy compounds with variations at the 14-position and the butenolide ring were much less active than the corresponding 3beta-hydroxy analogs. Thus the activity of 3-deoxydigitoxigenin appears to be particularly susceptible to structural changes elsewhere in the molecule.
The synthesis of a 3beta-thiocyanatocardenolide is described. The compound exhibited about 0.1 times the cardiotonic effect of digitoxyigenin in the isolated frog heart preparation. At a dosage of 20 mg/kg in the intact rat, it elicited ECG changes similar to those seen with a 10-mg/kg dose of digitoxigenin. Studies also revealed the new cardenolide to be a reversible inhibitor of sodium- and potassium-activated adenosine triphosphatase.
An alcoholic extract of the seeds of Coronilla varia L. showed inhibitory activity against KB cells in culture and was fractionated through a series of partitions, column chromatography, and preparative layer chromatography to yield hyrcanoside, daphnoretin, scopoletin, and umbelliferone. Hyrcanoside was also tested in the PS mouse leukemia assay and showed borderline activity.
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The binding of digitoxigenin to lecithin model membranes was investigated by application of electron spin- and nuclear magnetic resonance methods. A digitoxigenin spin-label derivative was found to bind specifically to the pseudohexagonal rigid lattice of lecithin membranes. The binding is accomplished by some of the structural features which are required for the biological activity of cardiac glycosides. Digitoxigenin has a procooperative effect on dipalmitoyl-lecithin membranes. The induction of lipid phase separation is discussed as a hypothetic molecular mechanism of action of cardiac glycosides on biological membranes.
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