Correlation between the negative inotropic potency and binding parameters of 1,4-dihydropyridine and phenylalkylamine calcium channel blockers in cat heart

Summary Partially purified plasma membranes were prepared from cat ventricle. The purification factors for the calcium channel ligands (+)-³H-PN 200-110, ³H-nimodipine (1,4-dihydropyridines) and (–)-³-H-desmethoxyverapamil (a phenylalkylamine) were 3.1-, 3.4- and 2.9-fold, respectively, whilst -adrenoceptors labelled with (–)-³H-dihydroalprenolol were purified 3.0-fold.(+)-³H-PN 200-110 bound to 930±140 fmol/mg of membrane protein with a dissociation constant of 70 pmol/l at 25°C. Under the same conditions ³H-nimodipine bound to 490±24 fmol/mg of sites with a K _D of 120 pmol/l. (–)-³-H-desmethoxyverapamil bound to 530±55 fmol/mg of sites with a K _D of 2.47 nmol/l.Twelve 1,4-dihydropyridines were evaluated for binding inhibition constants (K _i) with (+)-³H-PN 200-110 and 13 phenylalkylamines with (–)-³-H-desmethoxyverapamil in radioligand binding assays. Of the twelve 1,4-dihydropyridines evaluated (±)-nitrendipine was the most potent with a K _i-value of 280 pmol/l, nifedipine had a K _i-value of 500 pmol/l and the weakest drug tested, (±)-Bay b4328, had a K _i-value of 14.3 nmol/l. The EC₅₀-values of the same 1,4-dihydropyridines to inhibit the electrically driven cat papillary muscle were 77- to 3,450-fold higher and little correlation existed between K _i and EC₅₀-values.Thirteen phenylakylamines were tested for their potency to inhibit (–)-³-H-desmethoxyverapamil binding. The most potent phenylalkylamine with respect to negative inotropy was (±)-D 595 with an EC₅₀-value of 794 nmol/l, the least potent substance was (±)-Sz 45 with an EC₅₀-value of 39.8 mol/l. The binding inhibition constants for the phenylalkylamines were 13-to 322-fold lower than the values for negative inotropy, but a significant positive correlation between the K _i and EC₅₀-values (n=12, r=0.84) was observed. The absolute differences may reflect the state-dependent binding of phenylalkylamines to the channel.QSAR analysis revealed nearly identical correlations between physicochemical substituent properties on the one hand and binding affinities or functional potency on the other hand. In both cases the electronic properties (F-constant) of ring substituents mainly determine the variance in potency.     

Second-generation K(ATP) channel openers

This review discusses structural aspects of second-generation K(ATP) channel openers (KCOs), which exhibit improved tissue-selectivity. Their therapeutic profile is debated with main focus on cardiac ischemia, asthma, and urinary incontinence.     

KATP channel openers: structure-activity relationships and therapeutic potential

ATP-sensitive potassium channels (K(ATP) channels) are heteromeric complexes of pore-forming inwardly rectifying potassium channel subunits and regulatory sulfonylurea receptor subunits. K(ATP) channels were identified in a variety of tissues including muscle cells, pancreatic beta-cells, and various neurons. They are regulated by the intracellular ATP/ADP ratio; ATP induces channel inhibition and MgADP induces channel opening. Functionally, K(ATP) channels provide a means of linking the electrical activity of a cell to its metabolic state. Shortening of the cardiac action potential, smooth muscle relaxation, inhibition of both insulin secretion, and neurotransmitter release are mediated via K(ATP) channels. Given their many physiological functions, K(ATP) channels represent promising drug targets. Sulfonylureas like glibenclamide block K(ATP) channels; they are used in the therapy of type 2 diabetes. Openers of K(ATP) channels (KCOs), for example, relax smooth muscle and induce hypotension. KCOs are chemically heterogeneous and include as different classes as the benzopyrans, cyanoguanidines, thioformamides, thiadiazines, and pyridyl nitrates. Examples for new chemical entities more recently developed as KCOs include cyclobutenediones, dihydropyridine related structures, and tertiary carbinols.     

The importance of drug ionization for the action of calcium-antagonists and related compounds

pK-Values of Ca-antagonists and related cardiodepressive drugs have been measured by means of potentiometric microtitration. The presumable active species for all compounds investigated is the protonated molecule except nifedipine. This drug must exert its action via the uncharged form. From these results it could be concluded that protonization of ionizable nitrogen is one molecular prerequisite for voltage- or frequency dependence of action. Drug ionization does not correlate with the negative inotropic potency of the compounds investigated.     

Ca2+-Mg2+-ATPase activity of human red blood cells in healthy and diabetic volunteers

Summary The present investigation was dedicated to support biochemical interpretations of well-known long-term microvascular complications in diabetes. Provided the hypothetical correlation between erythrocyte membrane rigidity and increased intracellular calcium content holds true, a reduced Ca²⁺-Mg²⁺-ATPase activity in diabetic subjects could represent the underlying biochemical mechanism. Thus, we have compared basal and calmodulin-activated ATPase activity in healthy and diabetic volunteers. We could demonstrate a significant reduction of basal and stimulated enzyme activity in diabetic subjects. Furthermore, partial purification of calmodulin from erythrocytes of diabetic patients and healthy subjects yielded experimental evidence that reduced enzyme activity in diabetic volunteers is due to an altered basal activity as well as to a reduced stimulation by calmodulin.Abbreviations ATP Adenosine 5-triphosphate - ATPase Adenosine 5-triphosphatase - DEAE Diethylaminoethyl - EDTA Ethylenediamine tetraacetate - EGTA Ethyleneglycol-bis-(-aminoethyl ether)N,N-tetraacetic acid - P_i Inorganic phosphate - Tris Tris(hydroxymethyl)aminomethane     

Comparative QSAR studies on vasodilatory and negative inotropic properties of ring-varied verapamil congeners

In the present study quantitative structure-activity relationships (QSAR) for the vasodilator properties of verapamil have been derived: Potency of ring-varied verapamil congeners on the phasic and tonic component of K+-contractures has been measured in aortic rings of the cat. Potency of verapamil derivatives on the phasic component significantly correlates with lipophilic properties of the compounds. Inhibition of the tonic component best correlates with a parameter combination of steric and electronic properties (MV, sigma). These results contrast with investigations with dihydropyridine derivatives where identical SAR have been found for both response components. Furthermore, SAR analyses yield disparate results in comparison to investigations in heart muscle where primarily electronic and secondarily steric parameters best explain the potency of verapamil derivatives. The postulate of chemically differing binding sites for verapamil congeners in heart and smooth muscle is substantiated by calculating binding energies to the hypothetical binding site arginine. While negative inotropic potency of verapamil derivatives significantly correlates with binding energy to the model binding site arginine, this correlation fails in case of aortic ring. These results obtained for verapamil congeners contrast with observations with dihydropyridine derivatives where the chemistry of binding sites in heart and smooth muscle seems to be identical.