A simple and versatile method for the chemical synthesis of 21-hydroxypregnane 21-O-malonyl hemiesters which may be important intermediates of cardenolide biosynthesis is described. Starting from commercial beta-methyldigitoxin, acid hydrolysis followed by 3beta-O-acetylation and ozonolysis with reductive cleavage of the ozonides afforded 3beta-acetoxy-5beta-pregnane-14beta,21-diol-20-one which was finally converted into the target compound by treatment with malonyl chloride. The malonylation protocol was optimized using deoxycorticosterone (DOC) as the pregnane educt.