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The L-type calcium channel blocker nifedipine impairs extinction, but not reduced contingency effects, in mice.

We recently reported that fear extinction, a form of inhibitory learning, is selectively blocked by systemic administration of L-type voltage-gated calcium channel (LVGCC) antagonists, including nifedipine, in mice. We here replicate this finding and examine three reduced contingency effects after vehicle or nifedipine (40 mg/kg) administration. In the first experiment, contingency reduction was achieved by adding USs to the training protocol (degraded contingency), a phenomenon thought to be independent of behavioral inhibition. In the second experiment, contingency reduction was achieved by varying the percentage of CS-US pairing, a phenomenon thought to be weakly dependent on behavioral inhibition. In the third and fourth experiments, contingency reduction was achieved by adding CSs to the training protocol (partial reinforcement), a phenomenon thought to be completely dependent on behavioral inhibition. We found that none of these reduced contingency effects was impaired by nifedipine. In a final experiment, we found that extinction conducted 1 or 3 h post-acquisition, but not immediately, was LVGCC-dependent. Taken together, the results suggest that reduced contingency effects and extinction depend on different molecular mechanisms and that LVGCC dependence of behavioral inhibition develops with time after associative CS-US learning.

Animals↗

Effect of calcium channel blockers and trifluoperazine on rat liver regeneration.

The activity of hepatic thymidylate synthetase and thymidine kinase at 24 h after 70% partial hepatectomy of rats was suppressed significantly compared with that in the control group by the administration of calcium channel blockers (verapamil, diltiazem and nifedipine) 8 h after partial hepatectomy. The decrease of thymidylate synthetase and thymidine kinase activities was accompanied by a reduction of DNA content in 24 h regenerating liver. Trifluoperazine showed an effect similar to that of the calcium channel blockers on DNA synthesis during liver regeneration. These results suggest that calcium entry into the hepatic cell is an essential event in liver regeneration.

Animals↗

Particular sensitivity to calcium channel blockers of the fast inward voltage-dependent sodium current involved in the invasive properties of a metastastic breast cancer cell line.

1. A voltage-dependent sodium current has been described in the highly invasive breast cancer cell line MDA-MB-231. Its activity is associated with the invasive properties of the cells. The aim of our study was to test whether this current (I(Na)) is sensitive to three representative calcium channel blockers: verapamil, diltiazem and nifedipine. I(Na) was studied in patch-clamp conditions. 2. I(Na) was sensitive to verapamil (IC(50)=37.6+/-2.5 microM) and diltiazem (53.2+/-3.6 microM), while it was weakly sensitive to nifedipine. 3. The tetrodotoxin (TTX) concentration, which fully blocks I(Na) (30 microM), did not affect cell proliferation. Diltiazem and verapamil, at concentrations that do not fully block I(Na), strongly reduced cell proliferation, suggesting, regarding proliferation, that these molecules act on targets distinct from sodium channels. These targets are probably not other ionic channels, since the current measured at the end of a 500 ms long pulse in the voltage range between -60 and +40 mV was unaffected by verapamil and diltiazem. 4. We conclude that the sodium channel expressed in MDA-MB-231 cells is sensitive to several calcium channel blockers. The present study also underlines the danger of concluding to the possible involvement of membrane channel proteins in any phenomenon on the sole basis of pharmacology, and without an electrophysiological confirmation.

Breast Neoplasms↗

[Therapeutic value of antioxidants and calcium channel blockers in patients in the acute phase of closed head injuries].

Knowing that uncontrolled calcium signalization with excessive production of reactive oxidative matters is present in case of neurotrauma, aim of the investigation was to establish therapeutic value of combined administration of antioxidants (AO) and calcium channel blockers (CCB) in patients with closed head injury (CHI). Investigation comprised 120 patients with CHI who received AO (vitamins C and E) parenterally during 10 days and CCB (nimodipine), and control group was comprised of 60 patients with CHI who did not receive these medicinals in therapeutic program. We have established the influence of the therapy on neurologic and functional deficiency and consciousness disorder, respectively. Results of the investigation confirmed better recovery of all three observed parameters (degree of neurologic deficiency, degree of functional deficiency and consciousness disorder) in a group of patients receiving AO and CCB, which was statistically significant. It can be concluded that the administration of AO and CCB in patients with CHI in the acute phase should be included into therapeutic program of this significant clinical syndrome.

Adult↗

Antioxidant effects of calcium channel blockers against free radical injury in endothelial cells. Correlation of protection with preservation of glutathione levels.

The effects of four calcium channel blockers (nicardipine, nifedipine, verapamil, and diltiazem) on free radical injury in cultured endothelial cells were studied and compared with those of butylated hydroxytoluene. When the cultured cells were exposed to a superoxide and hydroxyl radical generating system for up to 60 minutes, lipid peroxidation occurred, and cellular viability decreased by 60% at 30 minutes. Concomitantly, total cellular glutathione decreased by 40%, whereas total protein thiols changed minimally. Preincubation of the cells with each of the calcium blockers (5 and 20 microM) before free radical addition resulted in various degrees of significant protection against cell death, and losses of glutathione correlated significantly (r = 0.89, p less than 0.001). The order of efficacy was nicardipine greater than nifedipine greater than verapamil greater than diltiazem; butylated hydroxytoluene was about fourfold more potent than nicardipine. Because none of the agents affected the level of hydroxyl radicals generated in the aqueous phase, the data suggest that the protective mechanisms were mediated by their lipid antiperoxidative activities, which also prevented the glutathione decrease caused by inhibition of peroxide generation.

Animals↗

Protective effect of labedipinedilol-A, a novel dihydropyridine-type calcium channel blocker, on myocardial apoptosis in ischemia-reperfusion injury.

The effects of labedipinedilol-A, a novel dihydropyridine-type calcium channel blocker with alpha-/beta-adrenoceptor blocking activities, on myocardial infarct size, apoptosis and necrosis in the rat after myocardial ischemia/reperfusion (45 min/120 min) were investigated. Ten minutes prior to left coronary artery occlusion, rats were treated with vehicle or labedipinedilol-A (0.25 or 0.5 mg/kg, i.v.). In the vehicle group, myocardial ischemia-reperfusion induced creatine kinase (CK) release and caused cardiomyocyte apoptosis, as evidenced by DNA ladder formation and terminal dUTP deoxynucleotidyltransferase nick end-labeling (TUNEL) staining. Treatment with labedipinedilol-A (0.25 or 0.5 mg/kg) reduced infarct size significantly compared to vehicle group (18.75+/-0.65% and 8.27+/-0.29% vs. 41.72+/-0.73%, P<0.01). Labedipinedilol-A also reduced the CK, CK-MB, lactate dehydrogenase (LDH) and troponin T levels in blood. In addition, labedipinedilol-A (0.5 mg/kg) significantly decreased TUNEL positive cells from 19.21+/-0.52% to 9.73+/-0.81% (P<0.01), which is consistent with absence of DNA ladders in the labedipinedilol-A group. Moreover, labedipinedilol-A pretreatment also decreased calcium content in ischemic-reperfused myocardial tissue. In conclusion, these results demonstrate that labedipindielol-A, through reduction of calcium overload and apoptosis, exerts anti-infarct effect during myocardial ischemia-reperfusion and would be useful clinically in the prevention of acute myocardial infarction.

Anesthesia↗

Comparison of inhibitory effects of calcium channel blockers and that of a calmodulin antagonist in strips of mesenteric arteries from spontaneously hypertensive and normotensive rats.

Effects of calcium channel blockers and of calmodulin antagonist on the contractile responses to norepinephrine (NE) were compared between strips of mesenteric arteries from 6- and 14-week-old spontaneously hypertensive rats (SHR) and age-matched, normotensive Wistar-Kyoto rats (WKY). The ratio of the maximum contraction developed by NE to that by 60 mM KCl was significantly increased in strips from 14-week-old SHR. Niludipine, verapamil and diltiazem antagonized the maximum NE contraction to a greater extent in strips from 14-week-old SHR than in those from the WKY. However, the antagonism by niludipine of the KCl- or caffeine-induced contraction was not significantly different between the strips from 14-week-old SHR and those from WKY. In strips from 6-week-old rats, there was no difference in the antagonism by niludipine of the maximum NE contraction. On the other hand, the effect of W-7 on the maximum NE contraction was not significantly different between the strips from 14-week-old SHR and those from WKY. Schild plot analyses demonstrated that alpha 1-adrenoceptors were the same for the strips from SHR and WKY. These results suggest that the enhanced maximum NE contraction in the mesenteric artery from 14-week-old SHR reflects the increased transmembrane influx of calcium, and the activity of calmodulin seems to be the same for the two strains.

Adrenergic alpha-Antagonists↗

Synthesis and biological activity of novel calcium channel blockers: 2,5-dihydro-4-methyl-2-phenyl-1,5-benzothiazepine-3-carboxylic acid esters and 2,5-dihydro-4-methyl-2-phenyl-1,5-benzodiazepine-3-carboxylic acid esters.

2,5-Dihydro-4-methyl-2-phenyl-1,5-benzothiazepine-3-carboxylic acid esters, based on the structures of dihydropyridines and diltiazem, were synthesized from o-aminothiophenol and 2-(phenylmethylene)- 3-oxobutanoic acid esters. Biological evaluation in the potassium-depolarized rabbit aorta suggests that these compounds are calcium channel blockers. The in vitro activity was further confirmed by electrophysiological techniques. Structure-activity studies for the aromatic substitution showed that the 2-nitro derivative was the most potent (IC50 = 0.3 microM) compound in vitro while the ethyl ester was slightly better than the corresponding methyl ester. Replacement of sulfur with nitrogen atom provided 2,5-dihydro-4-methyl-2-(3-nitrophenyl)-1,5-benzodiazepine-3-carboxylic acid ethyl ester, which was only slightly less active than the corresponding benzothiazepine. Derivatization of the nitrogen in 2,5-dihydro-4-methyl- 2-(3-nitrophenyl)-1,5-benzothiazepine-3-carboxylic acid methyl ester with a (dimethylamino)ethyl group (present in diltiazem) provided 2,5-dihydro-5-[(dimethylamino)ethyl]- 4-methyl-2-(3-nitrophenyl)-1,5-benzo-thiazepine-3-carboxylic acid methyl ester, which was found to be equipotent to diltiazem in vitro. Radioligand binding studies using [3H]nitrendipine and [3H]diltiazem showed that the compound with the free nitrogen binds competitively into the dihydropyridine binding site while the molecule in which the nitrogen is alkylated with a (dimethylamino)ethyl group interacts competitively with both diltiazem and dihydropyridine binding sites. Our results therefore show that 2,5-dihydro-4-methyl-2-phenyl-1,5-benzothiazepine-3-carboxylic ester is a good starting point for designing dihydropyridine as well as diltiazem mimics.

Action Potentials↗

Binding of the calcium channel blocker nitrendipine to its receptor in purified sarcolemma from canine cardiac ventricle.

High affinity binding of the 1,4-dihydropyridine calcium channel blocker [3H]nitrendipine was found in cardiac sarcolemma but not cardiac sarcoplasmic reticulum or mitochondria. Sarcolemmal binding of [3H]nitrendipine was saturable and reversible, with a maximum (Bmax) of approximately 1 pmol/mg protein and a Kd of approximately 0.14 nM. Displacement of sarcolemma-bound [3H]nitrendipine by other nifedipine analogs was stereospecific. The Kd for nitrendipine binding was approximately three orders of magnitude lower than the IC50 for the negative inotropic effect of this drug on isolated cat myocardium.

Animals↗

Role of calcium channel blockers in protection against experimental renal injury.

The available evidence indicates that the first generation calcium channel blocker verapamil has a protective effect against both acute and chronic renal failure. At the cell membrane, verapamil helps minimize the effects of excess calcium influx after ischemic injury, evidenced by reduced uptake of 45Ca, thus lessening tubular injury from both calcium-activated phospholipases and mitochondrial calcium overload. In experimental chronic renal failure, the long-term administration of verapamil protects against renal dysfunction and damage, independent of any effect on systemic mean arterial pressure. Protective effects of verapamil are delineated in several models, supporting the presented hypothesis of the pathogenesis of renal failure.

Acute Kidney Injury↗

[Exclusive use of calcium channel blockers and cardioselective beta-blockers in the pre- and per-operative management of pheochromocytomas. 70 cases].

The pre and intraoperative use of calcium channel blockers (CCB) has been suggested for the management of either eutopic or ectopic pheochromocytomas. We report our experience of 70 pheochromocytomas, operated between 1988 and 1996 and managed with CCB, especially nicardipine. 59 were hypertensive (84.2%). Preparation consisted of nicardipine in 61 patients or another CCB in 9 cases with duration ranging from 24 hours to several weeks depending on plasma volume and blood pressure control. Intraoperatively, nicardipine infusion was started after intubation, adjusted according to systolic blood pressure (SBP) and stopped before ligation of the tumor venous drainage. Increases in SBP greater than 200 mmHg were observed in 10 patients and were effectively controlled by nicardipine in all cases. In 16 patients, the S > BP remained less than 150 mmHg throughout anesthesia. Heart rate greater than 100 b p m occureed in 51 patients and was easily controlled with esmolol whenever used (n = 27). Arythmias were unfrequent (n = 4) and required treatment in only one case. This study confirms the ability to adequately manage pheochromocytomas with the use of nicardipine as sole vasodilating agent.

Adolescent↗

[The effect of the subchronic administration of calcium-channel blockers on CNS excitability].

The effect of a single day administration of nifedipine (10 mg/kg), cinnarizine (25 mg/kg), verapamile (25 mg/kg), fendiline (10 mg/kg), dilthiazem (10 mg/kg), and diazepam (0.5 mg/kg) on the dynamics of changing a convulsive dose of convulsant was studied on a model of intermittent intracutaneous administration of Corazole (10 mg/kg until convulsions) during 5 days. Repeated intermittent Corazole administrations were found to increase the sensitivity of animals with respect to convulsive effect of the drug. All calcium channel blockers and diazepam increased the convulsive dose of Corazole. In accordance with the hypothesis of anxiety disturbances based on nonspecific CNS activation, a possibility of application of calcium channel blockers as potential anxiolytics is considered.

Animals↗

Mechanical and thermal antinociception in rats following systemic administration of mibefradil, a T-type calcium channel blocker.

Voltage-gated Ca(2+) channels play a crucial role in pain perception. We studied the antinociceptive potential of systemically administered mibefradil, a peripherally acting antihypertensive agent and preferential T-type Ca(2+) channel blocker. Systemic injections of mibefradil, in clinically relevant doses, induced mechanical and thermal antinociception in adult rats without compromising their sensorimotor abilities. This suggests that peripherally acting T-type Ca(2+) channel blockers can be useful antinociceptive agents.

Analgesics↗

Effects of voltage-sensitive calcium channel blockers on extracellular dopamine levels in rat striatum.

Various subtypes of voltage-sensitive calcium channels (VSCCs) support the release of dopamine (DA) in the central nervous system. Using in vivo microdialysis, we investigate the influence of these subtypes of calcium channels on dopaminergic terminals in the rat striatum. L-type (nifedipine-sensitive), N-type (omega-conotoxin GVIA-sensitive), or N- and P/Q-type (omega-conotoxin MVIIC-sensitive) Ca2+ channels were blocked using selective antagonists injected locally, and K+-evoked DA release was measured in freely moving animals. K+ (100 mM) induced a massive increase of basal DA extracellular levels (930%) and was without significant effect on extracellular levels of DA metabolites DOPAC and HVA, and on the serotonin metabolite 5HIAA. Omega-conotoxin GVIA (1 microM) and omega-conotoxin MVIIC (1 microM) significantly reduced the K+-evoked DA release by 55 and 62%, respectively. The simultaneous application of the two conotoxins at the same concentration reduced K+-evoked DA release by 66%. Nifedipine (10 microM) had no significant effect on K-evoked DA release, while neomycin, a nonspecific VSCC blocker, produced a highly significant decrease when applied at 250 and 500 microM (56 and 75%, respectively). The compounds. however, had no effect on basal DA release and on the levels of extracellular DOPAC, HVA, and 5HIAA. These results suggest that under high and persistent conditions of membrane depolarization (15 min, 10 mM K+), striatal DA release is mainly mediated by N-type VSCCs.

3,4-Dihydroxyphenylacetic Acid↗

[Effects of NC-1100, a calcium channel blocker, on experimental cerebral ischemia/anoxia in rodents].

The anti-ischemic and anoxic effects of NC-1100, a piperazine type calcium channel blocker, were investigated in various cerebral ischemia and anoxia models in mice, gerbils and guinea pigs. Minimal effective doses of NC-1100 were 8 mg/kg, i.p. and 30 mg/kg, p.o. for KCN-induced anoxia; 16 mg/kg, i.p. for decapitation-induced gasping; 30 mg/kg, i.p. for cerebral ischemia induced by occlusion of bilateral carotid arteries in gerbils; and 10 microM for the in vitro ischemic model in hippocampal slices. Moreover, NC-1100 attenuated the disturbance of cerebral energy metabolism induced by decapitation in mice. These results suggest that NC-1100 has a cerebral protective effect, and that attributable to its ability to improve the cerebral energy metabolism disturbance.

Animals↗

In vitro effects of calcium channel blockers and beta-adrenergic blocking agents on microsomal lipid perocidation and cytochrome p-450 content.

The effects of the calcium channel blockers (CCB) nifedipine (N), verapamil (V) and diltiazem (D) and the beta adrenergic blocking agents (BAB) propranolol (P) and atenolol (A) administered alone or in combination on lipid peroxidation (LPO) and cytochrome p-450 content were studied in rat liver microsomes. The drugs were tested in concentrations of 1 mM. V, A and P alone significantly decreased TBARS formed after in vitro stimulation of LPO by Fe2+ and ascorbate, whereas no antioxidant effect was found for N and D. A correlation between the antioxidant capacity of the drugs and their ability to protect cytochrome p-450 after in vitro stimulation of LPO was observed except for propranolol. Moreover, propranolol abolished cytochrome p-450 protecting effect of verapamil when administered together. A direct, LPO-independent decreasing effect on cytochrome p-450 was observed upon in vitro incubation of microsomes with propranolol. The results are discussed in terms of LPO-dependent degradation of cytochrome p-450, formation of propranolol reactive metabolites and propranolol-dependent changes in cytochrome lipid environment.

Adrenergic beta-Antagonists↗

Influence of certain calcium-channel blockers on some aspects of lorazepam-dependence in mice.

The effect of acute and chronic treatments of the calcium-channel blockers, isradipine, diltiazem and flunarizine in protecting against lorazepam dependence has been demonstrated in mice. Dependence was induced by twice-daily administration of lorazepam (1 mg kg-1) for 10 days, doubling the dose during the next 10 days. Withdrawal symptoms and changes in the noradrenaline, dopamine and 5-hydroxytryptamine content of different regions of the brain were observed after either 24-h withdrawal or flumazenil administration. Isradipine inhibited lorazepam withdrawal symptoms, the effect being accompanied in the 24-h withdrawal group by significant decreases in the noradrenaline and dopamine content of the thalamus and hypothalamus and in the noradrenaline content of the mid-brain. In the flumazenil-treated group isradipine produced significant decreases in mid-brain noradrenaline and dopamine levels and in the dopamine content of the thalamus and hypothalamus. Diltiazem did not, on the other hand, afford significant protection against lorazepam withdrawal symptoms and did not induce any significant change in the neurotransmitters studied. Flunarizine significantly inhibited lorazepam withdrawal symptoms, an effect accompanied by significant reduction in noradrenaline and dopamine levels in the thalamus and hypothalamus. Dopamine was also significantly reduced in the cerebral cortex. Similar effects were produced in the flumazenil-treated group, and the noradrenaline content was reduced in the medulla, pons and cerebellum. It was concluded that isradipine and flunarizine might be of value in ameliorating lorazepam withdrawal symptoms.

Animals↗