Biomedical subjects

M Spedding

Publications and source records attributed to M Spedding.

At least 91 records · Page 5Linked to original sources

PubMed1988

Cerebral ischaemia reduces the density of 5-HT2 binding sites in the frontal cortex of the gerbil.

The 5-HT2 antagonist [3H]ketanserin labels a single population of high affinity sites (Kd 0.48 +/- 0.03 nM; Bmax 206 +/- 20 fmol/mg protein) in the frontal cortex of the gerbil. Specific binding of [3H]ketanserin was displaced by a number of 5-HT2A antagonists ritanserin, cyproheptadine and methysergide) but not by the 5-HT1A agonist, 8-hydroxy-2-(di-n- propylamino)tetralin (8-OH-DPAT) or the 5-HT1A/1B agonists 5-carboxyamidotryptamine (5-CT) or RU 24969, indicating that the labelled site probably represents the 5-HT2 receptor. Cerebral ischaemia induced in either a 3 hr unilateral non-recovery model or a 5 min bilateral, 3-day recovery model, resulted in a significant decrease in the density of 5-HT2 binding sites in the ischaemic frontal cortex without an apparent change in their affinity for the ligand. The decrease in density was not simply related to levels of 5-HT because occlusion of the right carotid artery for 3 hr resulted in bilateral depletion of 5-HT but only in an ipsilateral reduction in the density of binding sites. In addition, a significant decrease in the density of 5-HT2 binding sites occurred in the recovery model at a time when the levels of 5-HT in the cortex were unaltered.

Animals↗

PubMed1988

Antagonists and activators at calcium channels. Effects in the gastrointestinal tract.

Most calcium antagonists have relatively minor effects in the gastrointestinal (GI) tract; the factors influencing drug selectivity are reviewed. Voltage-dependent Ca channels (VOCs) in GI smooth muscle appear to be essentially similar to those in the cardiovascular system in their sensitivity to calcium antagonists. However, selective conditions for channel activation may allow antagonists to be selective in certain disease conditions. Indirect effects on blood flow may be crucial to certain parameters (e.g., acid secretion). Activators of VOCs have similar effects in vascular and GI smooth muscle. Some endogenous activators of VOCs may exist and evidence for control of VOC activity by lipid metabolites is indicated. Acyl carnitines may function as endogenous activators of VOCs.

Animals↗

PubMed1987

Calcium antagonist properties of diclofurime isomers. I. Functional aspects.

Trans-diclofurime has been shown to be a potent calcium antagonist which resembles verapamil in vitro and in vivo. Trans-diclofurime was a potent antagonist of Ca2+-induced contractions in K+-depolarized taenia preparations from the guinea pig caecum (pA2 = 8.3 +/- 0.2), whereas the cis isomer was 50 times less active (pA2 = 6.6 +/- 0.1); the inhibitory effects of trans-diclofurime were reversed noncompetitively by the Ca2+ channel activator Bay K 8644. Trans-diclofurime and verapamil were equipotent inhibitors of electrically evoked contractions of guinea pig left atria preparations; the inhibitory effects were frequency dependent and cis-diclofurime was 10 times less effective. Both diclofurime isomers prolonged the effective refractory period at high concentrations, indicating that they also possess local anaesthetic properties. Trans-diclofurime and verapamil reduced blood pressure in pithed rats infused with angiotensin II. Hypotensive effects were accompanied by bradycardia and prolongation of PR intervals, leading to second-degree atrioventricular block. The cis isomer was less potent. Diclofurime is thus a very potent calcium antagonist in heart and smooth muscle and has some additional membrane-stabilizing properties.

Animals↗

PubMed1987

Calcium antagonist properties of diclofurime isomers. II. Molecular aspects: allosteric interactions with dihydropyridine recognition sites.

Trans-diclofurime has been shown to be a very potent class II calcium antagonist (see preceding report), and we have examined its molecular interactions with the different receptor sites at the Ca2+ channel. Trans-diclofurime did not affect [3H]nitrendipine binding to rat cortical membranes at 37 degrees C and showed weak inhibitory effects at 25 degrees C, whereas at 0 degrees C 80% of the binding was inhibited noncompetitively (IC50, 13 nM); cis-diclofurime was 22-fold less potent. Trans-diclofurime, like diltiazem, blocked the inhibitory effects of verapamil on [3H]nitrendipine binding. Trans-diclofurime is a potent displacer of [3H]diltiazem binding (IC50, 15 nM; IC50 for diltiazem, 55 nM); the diclofurime isomers showed high stereoselectivity, with high Hill coefficients (0.85-1.0). In contrast, the stereoselectivity of the isomers was lower as inhibitors of [3H]verapamil binding, as were the Hill coefficients (0.55-0.65). It is proposed that the functional potency of the diclofurime isomers as calcium antagonists can be explained on the basis of their relative affinities for the diltiazem site and that this site is coupled to the dihydropyridine site in a positive heterotropic allosteric manner. A model for the interaction of group II calcium antagonists with the Ca2+ channel is proposed.

Animals↗

PubMed1987

MDL 72567, a dihydropyridine calcium-antagonist, that causes vasodilation and direct sinus bradycardia.

MDL 72567 (2,6 dimethyl,3 methoxycarbonyl,4-(2-nitrophenyl), 5-(2-furoyl)1,4 dihydropyridine) was a potent antagonist of Ca2+-induced contractions in K+-depolarized taenia preparations from the guinea pig caecum (pA2 8.8 +/- 0.1). MDL 72567 was a potent displacer of [3H]nitrendipine binding from rat cortical membrane preparations (Ki 3.99 nM), indicating an effect at the dihydropyridine binding site, which is consistent with the finding that the inhibitory effects of MDL 72567 in smooth muscle were prevented by the dihydropyridine Ca2+ channel activator Bay K 8644. MDL 72567 slowed spontaneously beating rat atria preparations to a greater extent than did nifedipine, however, for a given negative inotropic effect. Furthermore, in pithed rat preparations infused with angiotensin II to elevate blood pressure, the hypotensive effects of MDL 72567 (3 nmol/kg-3 mumol/kg, intravenously, i.v.) were accompanied by bradycardia, whereas nifedipine, PY 108-068, and nicardipine lowered blood pressure without affecting heart rate. When compared with nifedipine, MDL 72567 caused less reflex tachycardia for a given fall in blood pressure, in anesthetized beagles and in conscious renal hypertensive dogs. In anesthetized dogs, MDL 72567 increased cardiac contractility at all hypotensive doses tested (30-3,000 nmol/kg, i.v.), whereas nifedipine caused profound myocardial depression at higher doses (1,000-3,000 nmol/kg, i.v.) even though the compounds had equivalent vasodilator effects. Thus, although MDL 72567 appears to cause a direct myocardial slowing that can partially offset reflex tachycardia, the compound has negligible negative inotropic effects and may therefore be useful in angina pectoris or even in congestive heart failure.

Animals↗

PubMed1987

Comparison between the effects of MDL 72567 and nifedipine on various cardiovascular parameters in conscious sinoaortic-denervated rats and sham-operated controls.

In this study the effects of a new calcium entry blocking agent, 2,6-dimethyl-3-methoxycarbonyl-4-(2-nitrophenyl)-5-(2-furoyl)-1, 4-dihydropyridine (MDL 72567), were compared with those of nifedipine on blood pressure, heart rate, ECG, and cardiac contractility indices in conscious sinoaortic baroreceptor-denervated (SA-denervated) rats and their sham-operated controls. In sham-operated rats, the calcium-entry blocking agents (0.1-2 mg/kg i.v.) produced equivalent falls in blood pressure. However, nifedipine caused a much greater reflex tachycardia which was accompanied by a negative inotropic effect, and with the highest dose (2 mg/kg), a prolongation of the PQ interval. MDL 72567 induced an increase in myocardial contractility. In SA-denervated rats, both drugs produced an enhanced fall in blood pressure accompanied by a negative inotropic effect. Nifedipine did not change heart rate in SA-denervated rats, whereas MDL 72567 caused bradycardia. Thus, in these experiments, MDL 72567 caused less reflex tachycardia for a given fall in blood pressure than nifedipine and was less likely to cause myocardial depression. These effects of MDL 72567 may represent valuable, clinically relevant advantages over nifedipine.

Animals↗

PubMed1987

Interaction of phorbol esters with Ca2+ channels in smooth muscle.

The phorbol ester 12-O-tetradecanoyl phorbol-13-acetate (TPA), a selective activator of protein kinase C, had no effect on the sensitivity to Ca2+ or verapamil of K+-depolarized taenia preparations from the guinea-pig caecum, despite the use of high concentrations (1 microM for 3 h); this preparation is sensitive to Ca2+ channel activators and antagonists. TPA (0.03-3 microM) caused a slow contraction of rat aorta preparations; the contractions were resistant to the calcium-antagonists nifedipine (0.01 microM), verapamil (10 microM), diltiazem (10 microM) and cinnarizine (10 microM), but were antagonized by N-(6-aminohexyl)-5-chloro-1-naphthalensulphonamide (W-7, 50-200 microM). Prolonged exposure to TPA (greater than 2 h) resulted in spontaneous contractions which were sensitive to verapamil (1 microM). Isoprenaline and sodium nitroprusside relaxed phenylephrine-induced contractions in rat aorta preparations. TPA (0.3 microM) blocked the maximal response to isoprenaline but not to sodium nitroprusside indicating that TPA did selectively activate protein kinase C under these experimental conditions. These findings indicate that protein kinase C activation does not result in direct effects on Ca2+ channel function, but may exert effects indirectly (e.g. by modifying intracellular sensitivity to Ca2+, Ca2+ extrusion, or cellular depolarization).

Animals↗

PubMed1987

Direct activation of Ca2+ channels by palmitoyl carnitine, a putative endogenous ligand.

1 Palmitoyl carnitine, a lipid metabolite which accumulates in cytoplasmic membranes during ischaemia, has been shown to resemble the Ca2+ channel activator, Bay K 8644, in K+-depolarized smooth muscle. Palmitoyl carnitine caused concentration-dependent (1-1000 mumol l-1) augmentations in the sensitivity to Ca2+ of K+-depolarized taenia preparations from the guinea-pig caecum. The (+/-)-isomer was equieffective with the (-)-isomer, whereas carnitine was ineffective and palmitic acid relaxed the tissues. The shift to the left of Ca2+ concentration-response curves induced by palmitoyl carnitine (100 mumol l-1) was additive with that of Bay K 8644 (1 mumol l-1). 2 The interactions of palmitoyl carnitine with the different classes of calcium-antagonist were similar to those seen with Bay K 8644. Schild plots of the calcium-antagonist effects of nifedipine were shifted to the right following preincubation of the taenia with palmitoyl carnitine (30-300 mumol l-1). The inhibitory effects of verapamil were especially sensitive to palmitoyl carnitine (100 mumol l-1). Whereas the potency of diltiazem as a calcium-antagonist was reduced by palmitoyl carnitine (100 mumol l-1), the inhibitory effects of the lipophilic class III calcium-antagonists, cinnarizine and flunarizine, were entirely resistant to palmitoyl carnitine (100 mumol l-1). 3 Although palmitoyl carnitine has detergent properties in high concentrations and lyses red blood cells, these effects were not Ca2+-dependent, nor were they modified by calcium-antagonists. Other detergents did not have selective interactions with Ca2+ channels. 4 Palmitoyl carnitine inhibited [3H]-nitrendipine, [3H]-verapamil and [3H]-diltiazem binding to rat cortical membranes with IC50 values (mumol l-1) of 120 +/- 1, 95 +/- 17 and 120 +/- 15 mumol l-1 respectively. The inhibition showed little temperature-dependence, in contrast to that of Bay K 8644, except for a small reduction in the IC50 value for [3H]-verapamil binding at 37 degrees C (42 +/- 5 mumol l-1). Palmitoyl carnitine interacted selectively with the Ca2+ channel, in that effects on ligand binding to alpha-adrenoceptors, beta-adrenoceptors and 5-HT1A receptors occurred only at 5-10 fold higher concentrations. 5 It is concluded that palmitoyl carnitine, at concentrations which have previously been shown to occur in the cytoplasm during myocardial ischaemia, may interact directly with Ca2+ channels and may therefore be considered as an endogenous modulator of channel function. The site of action differs from that of other agents.

Animals↗

PubMed1986

Quantitative analysis of the dihydropyridines, 3-(2-furoyl)-5-methoxycarbonyl-2,6-dimethyl-4-(2-nitrophenyl)- 1,4-dihydropyridine and nifedipine, by high-performance liquid chromatography with electrochemical detection.

An analytical method based on solvent extraction and reversed-phase high-performance liquid chromatographic separation with electrochemical detection has been developed for the dihydropyridines, 3-(2-furoyl)-5-methoxycarbonyl-2,6-dimethyl-4-(2-nitrophenyl) -1,4-dihydropyridine (MDL 72.567) and nifedipine. The analysis includes an internal standard of similar light sensitivity to correct for possible photodegradation during the procedure. The specificity of electrochemical detection precludes interference from oxidized metabolites. Total analysis time was 35 min per sample, and the detection limit for quantification was 1-2 ng/ml. Linear regression analysis gave calibration curves with coefficients or correlation of 0.9992 for MDL 72.567 (1-100 ng) and 0.997 for nifedipine (1-50 ng). Assays for within-run and day-to-day reproducibility gave coefficients of variation of 3.9% and 6.0%, respectively, at concentrations of 50 ng/ml. The method has been applied to the analysis of plasma levels of nifedipine and MDL 72.567 in dogs.

Animals↗

PubMed1985

Antagonism of Ca2+-induced contractions of K+-depolarized smooth muscle by local anaesthetics.

Drugs known to interact with Na+ channels were compared as antagonists of Ca2+-induced contractions of K+-depolarized taenia preparations from guinea-pig caecum. Tetracaine (apparent pA2 5.3 +/- 0.2), quinidine (5.2 +/- 0.1) quinine (5.1 +/- 0.1), d-propranolol (4.7 +/- 0.1), 1-propranolol (4.7 +/- 0.1), lignocaine (4.0 +/- 0.1) and procaine (3.6 +/- 0.1) displaced cumulative concentration-response curves to Ca2+ to the right without depression the maximal response. The slopes of Arunlakshana and Schild plots were close to unity for quinidine, quinine, lignocaine and procaine. These drugs relaxed the established Ca2+-induced contractions rapidly and thus the effects of these drugs resembled the effects of low concentrations of verapamil. However, the effects of the local anaesthetics were increased in the presence of sodium salicylate (5-10 mM) which increases the negative surface charge. In contrast the effects of verapamil were decreased by salicylate. Veratridine (10-100 microM), which activates Na+ channels, had only depressant effects on Ca2+-induced contractions. Thus, drugs acting on Na+ channels can also interact with Ca2+ channels but there are qualitative as well as quantitative differences between the effects of these drugs and those of drugs such as verapamil. These findings indicate different mechanisms of action for the inhibition of Ca2+-induced contractions by local anaesthetics and verapamil.

Anesthetics, Local↗

PubMed1985

Competitive interactions between Bay K 8644 and nifedipine in K+ depolarized smooth muscle: a passive role for Ca2+?

The kinetics of the interactions between Bay K 8644, a calcium channel activator, and the "calcium-antagonists" nifedipine, verapamil and diltiazem have been investigated. Nifedipine shifted cumulative concentration-response curves for Ca2+ to the right in K+ depolarized taenia preparations from the guinea-pig caecum. The apparent pA2 was 9.3 +/- 0.2 (slope 1.44; 95% confidence limits 0.99-1.88). Bay K 8644 (10-1,000 nmol/l) reduced the inhibitory effects of nifedipine, shifting the Schild plots to the right, without affecting the slope of the nifedipine:Ca2+ interaction. Thus, the interaction between the dihydropyridines was independent of external Ca2+. The parallel shifts of the Schild plots allow a novel interpretation of the "agonist" potency of Bay K 8644 because the compound had an apparent pA2 of 8.8 (slope 0.92) as an "antagonist" of the inhibitory effects of nifedipine. In contrast, Bay K 8644 was a non-competitive antagonist of the inhibitory effects of verapamil and diltiazem on Ca2+-induced contractions. These findings emphasize the differences between the various classes of "calcium-antagonists" and show that Bay K 8644 is a powerful tool discriminating between them.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

PubMed1985

Differential effects of calcium channel antagonists on histamine and pentagastrin-stimulated gastric acid secretion in the rat.

Different calcium channel antagonists have been assessed for their ability to inhibit gastric acid secretion in rats. Whereas verapamil, diltiazem and cinnarizine inhibited pentagastrin-induced gastric acid secretion but not histamine-induced secretion, nifedipine selectively inhibited the stimulant effect of histamine. In contrast, the vasodilator hydralazine had non-selective effects. These findings indicate that calcium-antagonists may have differential effects against different secretagogues and these effects are not simply related to hypotensive effects.

Anesthesia↗

PubMed1984

"Calcium antagonists": a class of drugs with a bright future. Part II. Determination of basic pharmacological properties.

This minireview discusses some simple pharmacological tests useful in detecting biological activity (screening), characterizing mechanisms of action and predicting possible therapeutic applications for calcium antagonists in general and calcium slow channel blockers in particular. In smooth muscle preparations these agents inhibit mechanical effects evoked by K+-depolarization which selectively opens voltage-operated calcium channels (VOC) to allow extracellular Ca++ into the cytosol. In contrast, any inhibition of receptor-mediated responses by calcium antagonists appears to depend on the transduction system and the specific cellular mechanism (e.g. VOC opening consequent to partial depolarization) activated by the receptor and, evidently, on ancillary pharmacological properties of the studied compound. For instance, whereas calcium slow channel blockers antagonize contractions produced by norepinephrine and K+-depolarization in the rat isolated portal vein, they inhibit effectively only the latter response in the rabbit aorta. This apparent discrepancy may be accounted for by the different pool of Ca++ mobilized in the two tissues by norepinephrine. Agents (e.g. diphenylalkylamines, calmodulin blockers) that impair the interaction of Ca++ with intracellular proteins produce effects which are less specific than those of slow channel blockers. Currently, the pharmacological profile of calcium antagonists can be appropriately defined by studying their effects on radioligand (dihydropyridine) binding, radioactive calcium movements through biological membranes, electrophysiological parameters in cardiac and vascular smooth muscle and on various in vivo cardiovascular preparations. Together, these approaches allow a functional classification of new calcium antagonists in relation to already known compounds and some hypotheses on their potential clinical applications. Finally, desirable pharmacokinetics and pharmacological properties for novel calcium antagonists are mentioned. This point will be further explored in the forthcoming minireview which will deal with the clinical applications of calcium antagonists.

Animals↗

PubMed1984

Interactions between a "calcium channel agonist", Bay K 8644, and calcium antagonists differentiate calcium antagonist subgroups in K+-depolarized smooth muscle.

The proposal that calcium antagonists have different sites of action has been tested by attempting to reverse their inhibitory effects with a dihydropyridine which augments Ca2+ entry into cells, Bay K 8644. Bay K 8644 (1-1000 nmol/l) increased the sensitivity to Ca2+ of K+-depolarized taenia preparations from the guinea-pig caecum. Thus Bay K 8644 augmented established submaximal Ca2+-induced contractions and also shifted cumulative concentration-response curves to Ca2+ to the left, even in the presence of an optimal K+-depolarization. The inhibitory effects of nifedipine (10 nmol/l), verapamil (0.2 mumol/l), diltiazem (1 mumol/l) and diclofurime (1 mumol/l) on Ca2+-induced contractions were reversed by Bay K 8644 (1-1000 nmol/l). In contrast, Bay K 8644 did not reverse the effects of cinnarizine (1 mumol/l), flunarizine (1 mumol/l), fendiline (3 mumol/l), prenylamine (3 mumol/l), pimozide (1 mumol/l), bepridil (3 mumol/l), perhexiline (10 mumol/l) or the calmodulin antagonist W-7 (200 mumol/l). Bay K 8644 (1-100 nmol/l) was less effective at reversing the effects of nisoldipine (10 nmol/l), a slowly dissociating dihydropyridine, than the effects of nifedipine. However, preincubation with Bay K 8644 (1 mumol/l) protected the taenia from the inhibitory effects of nisoldipine (10 nmol/l). These findings are compatible with interactions of nisoldipine and Bay K 8644 at a common site. Taenia preparations incubated with Bay K 8644 (1 mumol/l) were protected from the inhibitory effects of nifedipine (10 nmol/l), nisoldipine (10 nmol/l) and to a lesser extent verapamil (0.2 mumol/l) and diltiazem (1 mumol/l).(ABSTRACT TRUNCATED AT 250 WORDS)

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

PubMed1984

Changing surface charge with salicylate differentiates between subgroups of calcium-antagonists.

Sodium salicylate (5-10 mM) has been used to distinguish the effects of the three calcium-antagonist subgroups which had been previously differentiated in functional studies. Sodium salicylate (10 mM) reduced the antagonistic effects of verapamil and diltiazem on Ca2+-induced contractions of K+ (40 mM)-depolarized taenia preparations from the guinea-pig caecum. In contrast, salicylate had no effect on the potency of nifedipine and increased the inhibitory effects of cinnarizine and flunarizine. Sodium salicylate (10 mM) had little effect on Ca2+-induced contractions per se. In preparations pretreated with calcium-antagonists and recontracted with high concentrations of Ca2+, salicylate (5 mM) caused an additional contraction when the preparations had been pretreated with verapamil or diltiazem but had no effect in control or nifedipine-treated preparations. In contrast, salicylate relaxed Ca2+-induced contractions in tissues which had been pretreated with cinnarizine, flunarizine, pimozide, bepridil, fendiline, perhexiline and with the calmodulin antagonist W-7. The mechanism of action of salicylate was investigated. Inhibition of prostaglandin biosynthesis or of oxidative phosphorylation by salicylate was not responsible for these effects because indomethacin (28 microM) and 2,4-dinitrophenol (20 microM) did not differentiate between calcium antagonists. The effects of salicylate are ascribed to an increase in negative surface charge on the membrane because other agents changing surface charge (3,5-dichlorosalicylate, 0.3 mM; benzoate, 20 mM) have similar effects and their potency is dependent on their affinity for lipid membranes. Furthermore, salicylate increased the effectiveness of the cationic local anaesthetic, (+)-propranolol (100 microM), but did not change the effects of the neutral local anesthetic, benzocaine (1 mM). It is argued that salicylate increases the effectiveness of cinnarizine by increasing accumulation of this drug in the cell membrane or at intracellular sites whereas the reduced effectiveness of verapamil and diltiazem is secondary to a change in the state of the Ca2+ channel.

2,4-Dinitrophenol↗

PubMed1983

"Calcium antagonists": a class of drugs with a bright future. Part I. Cellular calcium homeostasis and calcium as a coupling messenger.

The aim of this series of minireviews is to present material from multidisciplinary sources to facilitate the understanding of the pharmacology and the ample clinical potential of a class of drugs that were originally designated as "calcium antagonists" and more recently have been referred to as "calcium entry blockers", "calcium slow channel blockers" or "calcium modulators". In this first report our attention will be focussed on the pivotal role of Ca++ as a messenger linking stimuli of extracellular origin to the intracellular environment. Eucaryotic cells have a number of powerful means to control their cytosolic Ca++ concentration. Firstly, in a cell at rest the cellular membrane is relatively impermeable to passive Ca++ movements. This property of the plasmalemma prevents the high free Ca++ concentration (approximately 1 mM) of the extracellular compartment from invading the cytosol (approximately 0.1 microM). However, extracellular Ca++ can reach the cytosol through the Na+/Ca++ exchange mechanism and the plasmalemma possesses special Ca++ channels the conductance of which is controlled by gates that are opened by critical changes in cellular polarization (voltage-operated channels: VOC) or by receptor activation (receptor-operated channel: ROC). The Ca++ entering via VOC or ROC can subsequently trigger the liberation of Ca++ from the sarcoplasmic reticulum or from calcium stores located in the inner side of the plasmalemma. The intracellular message generated by external stimuli is transferred to the response mechanism by several cytosolic proteins that require Ca++ as activator. Finally, the termination of the response is the result of a reduction in the cytosolic Ca++ concentration that is accomplished by the Na+/Ca++ exchange mechanism or by energy-dependent pumps which extrude Ca++ from the cell or store it in subcellular organelles. Therefore, any of the numerous steps of the excitation-response coupling which employ Ca++ as a messenger or as a protein activator can be the site of action of a pharmacological agent. In the follow-up minireview, some methods to determine the basic pharmacological profile of compounds interfering with cellular Ca++-dependent functions will be described.

Action Potentials↗

PubMed1983

Direct inhibitory effects of some 'calcium-antagonists' and trifluoperazine on the contractile proteins in smooth muscle.

1 Taenia preparations from the guinea-pig caecum were treated with Triton X-100 and glycerol to disrupt the plasma membrane. Disruption of the sarcolemma was confirmed by electronmicroscopy. The preparations contracted in response to low concentration of Ca2+ (10-40 microM) and the contractions were dependent upon exogenous adenosine triphosphate (ATP). 2 Nifedipine (100 microM), verapamil (100 microM) and diltiazem (100 microM) did not inhibit Ca2+-induced activation of the contractile proteins. 3 In contrast, fendiline (100 microM), cinnarizine (100 microM), flunarizine (100 microM), pimozide (100 microM) and trifluoperazine (100 microM) significantly inhibited Ca2+-induced contractions. The effects of cinnarizine (100 microM) were reversible. 4 These findings disclose further differences between calcium-antagonists and suggest that certain of these agents have an intracellular site of action.

Animals↗

PubMed1983

Functional interactions of calcium-antagonists in K+-depolarized smooth muscle.

The functional significance of the interaction of certain calcium-antagonists with nimodipine was examined so that this might be related to binding studies. To this end, the relaxant effects of nimodipine (1-100 nM) on Ca2+ (10 mM)-induced contractions of K+-depolarized taenia preparations from the guinea-pig caecum were compared in the presence of nifedipine and cinnarizine (which competitively displace [3H]-nimodipine with high and low affinities respectively), diltiazem (which increases binding), verapamil (which allosterically reduces binding) and W-7, a calmodulin antagonist. The relaxant effects of nimodipine were similar in the presence of nifedipine, diltiazem and cinnarizine, but were slightly attenuated in the presence of verapamil and W-7. These findings can be reconciled with the differentiation of calcium antagonists evident from [3H]-nimodipine binding studies, but indicate that the functional consequences of allosteric interactions disclosed in such studies are small. Drugs which bind to calcium channels and drugs which bind to calmodulin did not potentiate each other.

Animals↗