title: The two mechanisms of action of racemic cardiotonic EMD 53998, calcium sensitization and phosphodiesterase inhibition, reside in different enantiomers.
abstract: The novel cardiotonic EMD 53,998 increases contractile force in vitro through both inhibition of phosphodiesterase III (PDE III) activity and increase in the responsiveness of the contractile proteins to calcium ("calcium sensitization"). Because EMD 53,998 is a racemate, the possibility arose that the two modes of action do not reside equally in the enantiomers. Therefore, the effects of the racemate and its two enantiomers [(+)EMD 57,033 and (-)EMD 57,439] were analyzed in guinea pig and rat cardiac tissue with respect to Ca2+ sensitization (Ca(2+)-induced force development in skinned cardiac myofibers and myofibrillar ATPase activity) and PDE III inhibition (isolated PDE isoenzymes and cyclic AMP level in isolated cardiac myocytes). In addition, the positive inotropic effects were compared in isometrically contracting papillary muscles. Enhancement of force of contraction (Fc) in submaximally activated skinned fibers showed a selectivity for the (+)enantiomer with EC50 = 1.7, 4.8, and > 100 microM for EMD 57,033, EMD 53,998, and EMD 57,439, respectively. Ca2+ concentration for half-maximal activation was decreased by 0.5 log units, and Cmax was increased by 15% at 10 microM EMD 57,033. Similarly, myofibrillar ATPase activity was most potently enhanced by the (+)enantiomer, with EC50 values of 1.8, 2.5, and > 30 microM for EMD 57,033, EMD 53,998, and EMD 57,439, respectively. PDE III activity was inhibited with greater potency by the (-)enantiomer, with IC50 values of 0.05, 0.06, and 1.94 microM for EMD 57,439, EMD 53,998, and EMD 57,033, respectively. The cyclic AMP content of isoprenaline-stimulated rat cardiac myocytes was increased by 50% at 13.6 and 0.71 microM for EMD 57,033 and EMD 57,439, respectively. In intact guinea pig papillary muscle, the positive inotropic effect of the (+)enantiomer was insensitive to isoprenaline pretreatment; in contrast, the (-)enantiomer showed only a weak positive inotropic action which was strongly enhanced in the presence of isoprenaline. We conclude that one of the two different mechanisms underlying the overall positive inotropic activity of EMD 53,998 can be assigned, almost exclusively, to one of the two enantiomers. Thus, the (-)enantiomer EMD 57,439 is a "pure" PDE III inhibitor with almost no Ca2+ sensitizing activity; the (+)enantiomer EMD 57,033 is a potent Ca2+ sensitizer with only a weak PDE III inhibitory activity as compared with the racemate. In contrast to other compounds with mixed activity, EMD 57,033 is unique in possessing both a high absolute potency at the level of the contractile elements and a favorable relation of Ca2+ sensitization to PDE inhibition.
authors.affiliation: Pharmaceutical Research Department, E. Merck, Darmstadt, Germany.
authors.forename: I
authors.initials: I
authors.lastname: Lues
authors.forename: N
authors.initials: N
authors.lastname: Beier
authors.forename: R
authors.initials: R
authors.lastname: Jonas
authors.forename: M
authors.initials: M
authors.lastname: Klockow
authors.forename: G
authors.initials: G
authors.lastname: Haeusler
mesh_terms: D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002316:Cardiotonic Agents / Q000494:pharmacology*; D000242:Cyclic AMP / Q000494:pharmacology; D004558:Electric Stimulation; D006168:Guinea Pigs; D006321:Heart / Q000187:drug effects*; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000378:metabolism; D007537:Isometric Contraction / Q000187:drug effects; D008297:Male; D009200:Myocardial Contraction / Q000187:drug effects; D009210:Myofibrils / Q000201:enzymology; D010210:Papillary Muscles / Q000166:cytology / Q000187:drug effects; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*; D011804:Quinolines / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D013237:Stereoisomerism; D013829:Thiadiazines / Q000494:pharmacology*
chemical_list: D002316:Cardiotonic Agents; D007527:Isoenzymes; D010726:Phosphodiesterase Inhibitors; D011804:Quinolines; D013829:Thiadiazines; C070609:EMD 53998; D000242:Cyclic AMP; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D000251:Adenosine Triphosphatases; D002118:Calcium