Classes Of Calcium Channel Blockers Research Articles

Effect of Calcium Channel Blockers on Platelet GPIIb-IIIa as a Calcium Channel in Liposomes: Comparison with Effects on the Intact Platelet

The platelet membrane glycoprotein IIb-IIIa complex is essential for platelet aggregation and functions as a fibrinogen receptor on the activated platelet. When incorporated into phospholipid vesicles, this glycoprotein complex can function as an apparent calcium channel which facilitates the transit of calcium across a phospholipid barrier. In order to further evaluate this calcium channel, the effect of calcium channel blockers of the dihydropyridine (nifedipine and nicardipine), arylalkylamine (verapamil) and benzothiazepine (diltiazem) classes were evaluated on GPIIb-IIIa liposomes with encapsulated fura-2 (a fluorescent calcium indicator). Nicardipine, verapamil, and nifedipine significantly inhibited calcium influx into GPIIb-IIIa liposomes; however, this required 190 microM, 400 microM, and 140 microM drug, respectively. These concentrations are 10-1,000 fold greater than those clinically obtainable. In contrast, diltiazem at concentrations greater than 220 microM and amiloride at concentrations greater than 800 microM showed no inhibitory effects. When aspirinized platelets were activated with 30 micrograms/ml bovine fibrillar collagen, both nicardipine and diltiazem produced a decrease in both the initial rise and maximum cytoplasmic calcium concentration. Parallel experiments were performed to assess the effects of verapamil, nicardipine, and diltiazem on platelet aggregation in platelet rich plasma. Nicardipine, 190-380 microM, induced a prolongation of the lag phase, but no effect on the final degree of platelet aggregation to collagen. Similar inhibition of platelet aggregation was seen with diltiazem and verapamil although the effect of diltiazem was less pronounced particularly at higher concentrations of collagen. No effect was seen on aggregation with 32 microM ADP which is release independent, or on the primary wave of low dose ADP induced platelet aggregation.(ABSTRACT TRUNCATED AT 250 WORDS)

Calcium channel blockers and gingival overgrowth

Calcium channel blockers are widely used in medical practice for the management of cardiovascular disorders. Of concern to the dental practitioner is the effect of these drugs on the gingival tissues. All classes of calcium channel blockers have been implicated in causing gingival overgrowth. This review considers the prevalence, clinical features, histopathology, pathogenesis and management of this unwanted side-effect.

Voltage‐dependent calcium channels regulate GH4 pituitary cell proliferation at two stages of the cell cycle

Calcium is an intracellular signal implicated in the regulation of cell proliferation. We have examined the growth regulatory role of voltage-dependent calcium channels (VDCC) in a rat pituitary cell line (GH4C1) that expresses two well-characterized VDCC subtypes (L and T) and is growth-inhibited by several agents known to enhance calcium entry. Thyrotropin-releasing hormone (TRH), tetradecanoylphorbol acetate (TPA), and epidermal growth factor (EGF), each known to enhance calcium entry in GH4 cells, decrease GH4 cell number and incorporation of [3H]-thymidine. The growth inhibitory action of these agents is cytostatic with a predominant effect to block G1 cells from entering S-phase. We next examined the growth regulatory action of pharmacologic agents that interact directly and specifically with type L VDCC. Activation of type L VDCC with the dihydropyridine BAY K8644 inhibits GH4 proliferation as measured by cell number and [3H]-thymidine incorporation. This action of BAY K8644 is enhanced by a submaximal K(+)-maintained depolarization, and the growth inhibitory action of these agents is also cytostatic as evident by the block of G1 cells from entering S-phase. Nimodipine, an antagonist specific for type L VDCC blocks (IC50 = 30 nM) BAY K8644-inhibited cell proliferation by substantially reducing the S-phase block. Taken together these findings indicate that calcium entry through type L VDCC inhibits GH4 cell proliferation by blocking entry into S-phase. By contrast, nimodipine caused only a small reversal of the TRH-induced S-phase block, suggesting that TRH inhibits proliferation by a mechanism that differs at least in part from L-channel activation. Unexpectedly, nimodipine, given alone, caused a substantial inhibition of GH4 cell proliferation. This action of nimodipine was cytostatic, yet differed from calcium channel activators in that the percentage of S-phase cells was unchanged whereas G2-M-phase cells increased with a parallel decrease in G1-phase cells. Similar effects were also observed with other classes of calcium channel blockers. Taken together these results indicate that calcium entry through VDCC regulates GH4 cell proliferation differently depending on the stage of the cell cycle. In G1-phase cells, sustained entry of calcium through type L VDCC blocks entry into S-phase. In G2-M-phase cells entry of calcium promotes progression through mitosis.

Antagonistes calciques. I. Remplacement du groupe diméthoxy-3,4 phényléthyle du vérapamil

In the verapamil structure, the 3,4-dimethoxy phenylethyl group has been replaced by several moieties belonging to α- and β-blockers or other classes of calcium channel blockers. The best derivatives, as calcium blockers, were prepared with β-blocker fragments like aryloxypropanolamines (AOPA) with an ortho substituent on the aromatic ring.

Calcium channel agonists and antagonists: effects of chronic treatment on pituitary prolactin synthesis and intracellular calcium.

PRL synthesis by GH cells in culture has previously been shown to increase when calcium is added to cultures grown in calcium-depleted medium or when cultures are treated for 18 h or longer with the dihydropyridine calcium channel agonist BAY K8644, whereas the antagonist nimodipine inhibits PRL. The experiments described here were designed to test whether differences in PRL synthesis caused by the dihydropyridines are due to changes in PRL mRNA levels, whether structurally different classes of calcium channel blockers alter PRL production, and whether long term treatment with calcium channel agonists and antagonists alters intracellular free calcium, [Ca2+]i. PRL synthesis and PRL mRNA levels were increased similarly by BAY K8644 and decreased in parallel by the dihydropyridine antagonist nimodipine, while overall protein and RNA synthesis were not changed by either the agonist or antagonist. Two calcium channel blockers which act at different sites on L-type channels than the dihydropyridines also inhibited PRL synthesis without affecting GH; 5 microM verapamil reduced PRL by 64% and 15 microM diltiazem by 89%. Partial depolarization with 5-25 mM KCl increased PRL synthesis up to 2-fold. The intracellular free calcium ion concentration was estimated by Quin 2 and averaged 142 nM for control cultures in normal medium, and 128 and 168 nM for cultures treated 72 h with nimodipine or BAY K8644, respectively. Nimodipine totally prevented the calcium rise obtained upon depolarization.(ABSTRACT TRUNCATED AT 250 WORDS)

Interaction between propanolol and calcium channel blockers in cardiac and vascular smooth muscle

The ability of the beta-receptor antagonist propranolol to influence the response of isolated cardiac and vascular smooth muscle to several classes of calcium channel blockers was examined. For comparison, the interactions between propranolol and other classes of negative inotropic and vasorelaxant agents was also evaluated. The results of these studies demonstrate that propranolol pre-treatment significantly enhances the in vitro response to the dihydropyridine calcium channel blocker nifedipine, but not the thiazapine calcium channel blocker diltiazem. This enhancement was unrelated to the negative inotropic or vasorelaxant properties of these agents. In addition, propranolol pre-treatment of rat cortical membranes also enhanced the affinity of nifedipine for the 3H-nitrendipine binding site, but did not alter the effect of diltiazem on 3H-nitrendipine binding. These observations suggest that a direct interaction may exist between beta-receptor antagonists and dihydropyrine-type calcium channel blockers. This interaction may be an important factor in selecting drug therapy for conditions such as hypertension and angina.

Identification of the multidrug resistance-related membrane glycoprotein as an acceptor for calcium channel blockers.

A radioactive photoactive dihydropyridine calcium channel blocker, [3H]azidopine, was used to photoaffinity label plasma membranes of multidrug-resistant Chinese hamster lung cells selected for resistance to vincristine (DC-3F/VCRd-5L) or actinomycin D (DC-3F/ADX). Sodium dodecyl sulfate-polyacrylamide gel electrophoretic fluorograms revealed the presence of an intensely radiolabeled 150-180-kDa doublet in the membranes from drug-resistant but not from the drug-sensitive parental (DC-3F) cells. A similar radiolabeled doublet was barely detected in a drug-sensitive partial revertant (DC-3F/ADX-U) cell line. The 150-180-kDa doublet exhibited a specific half-maximal saturable photolabeling at 1.07 X 10(-7) M [3H]azidopine. The dihydropyridine binding specificity was established by competitive blocking of specific photolabeling with nonradioactive azidopine as well as with nonphotoactive calcium channel blockers nimodipine, nitrendipine, and nifedipine. In addition, [3H]azidopine photolabeling was blocked by verapamil and diltiazem but was stimulated by excess prenylamine and bepridil suggesting a cross-specificity for up to four different classes of calcium channel blockers. The 150-180-kDa calcium channel blocker acceptor co-electrophoresed exactly with the 150-180-kDa surface membrane glycoprotein (gp150-180 or P-glycoprotein) Vinca alkaloid acceptor from multidrug-resistant cells and was immunoprecipitated by polyclonal antibody recognizing gp150-180. [3H]Azidopine photolabeling of the 150-180-kDa component in the presence of excess vinblastine was reduced over 90%, confirming the identity or close relationship of the calcium channel blocker acceptor and the gp150-180 Vinca alkaloid acceptor. The [3H]azidopine photolabeling of gp150-180 also was reduced by excess actinomycin D, adriamycin, or colchicine, demonstrating a broad gp150-180 drug recognition capacity. The ability of gp150-180 to recognize multiple natural product cytotoxic drugs as well as calcium channel blockers suggests a direct function for gp150-180 in the multidrug resistance phenomenon and a role in the circumvention of that resistance by calcium channel blockers.

Current therapy, present limitations and future goals for systemic hypertension

During the past 2 decades, encouraging strides have been made in the recognition and treatment of patients with high blood pressure and in the development of antihypertensive therapy. Concerns surrounding the control of hypertension, however, continue to emerge. As many as two-thirds of hypertensive patients in the US remain inadequately controlled. Although overall cardiovascular mortality has declined since the mid-1960s, a significant impact of antihypertensive therapy on coronary artery disease morbidity and mortality has not been conclusively demonstrated. Unfavorable metabolic effects associated with traditional step-care antihypertensive therapy with diuretics and β blockers are increasingly implicated for failure to demonstrate a decrease in coronary artery disease. The availability of newer efficacious antihypertensive therapy that does not adversely affect metabolic parameters is resulting in a reappraisal of traditional step-care. Of particular interest are the calcium channel blockers, because they are efficacious, well tolerated and offer favorable metabolic and hemodynamic profiles. Nitrendipine is a new long-acting member of the 1,4 dihydropyridine class of calcium channel blockers that has shown encouraging results for treatment of hypertension.

Diltiazem-like effect of thioridazine on the dihydropyridine binding site of the calcium channel of rat myocardial membranes

The phenothiazine thioridazine enhances the binding of [ 3H]-nitrendipine to cardiac membranes and antagonizes the inhibition of [ 3H]-nitrendipine binding by other classes of calcium channel blockers. These effects are similar to those of diltiazem. The increased binding of [ 3H]-nitrendipine in the presence of thioridazine or diltiazem is due to increased binding affinity rather than an increased number of binding sites. Increased affinity is due in part to a decrease in the dissociation rate of nitrendipine in the presence of either thioridazine or diltiazem. Thioridazine also reverses the blocking effect of tiapamil (a phenylalkylamine) on [ 3H]-nitrendipine binding to rat heart membranes, as does diltiazem, suggesting that thioridazine binds to the same site as diltiazem. Thioridazine may have calcium channel blocking effects in the high therapeutic and toxic dose ranges that could be responsible for some of its adverse cardiovascular effects.

Solubilization and partial purification of putative calcium channels labelled with [3H]-nimodipine.

High-affinity binding sites for the potent 1,4-dihydropyridine calcium channel blocker [3H]-nimodipine were solubilized from guinea-pig skeletal muscle microsomes with digitonin and CHAPS [3-(3-cholamidopropyl)-dimethyl-ammonio-l-propanesulfonate]. Detergent-solubilized binding sites could not be sedimented by centrifugation (50,000 X g, 4 h), passed freely through 0.2 micron nitrocellulose filters and were stable at 4 degrees C with half-lives of greater than 60 h. The solubilized 1,4-dihydropyridine binding sites were precipitable with polyethyleneglycol 6000 on Whatman GF/C filters. Saturation analysis of solubilized microsomes with [3H]-nimodipine revealed a single class of binding sites (Bmax = 0.5 to 1.7 pmol per mg of protein) with a KD of 2.2-3.6 nmol/l at 37 degrees C. Specific binding of the 1,4-dihydropyridine calcium channel label was fully reversible (k-1 = 1.5 min-1, at 37 degrees C). The solubilized drug receptors discriminated between the optical enantiomers of chiral 1,4-dihydropyridine calcium channel blockers, (-)- and (+)D-600 as well as between l-cis and d-cis-diltiazem. d-cis-Diltiazem stimulated the binding of [3H]-nimodipine (ED50:3.6 mumol/l), by increasing the Bmax and slowed the dissociation rate of the labelled 1,4-dihydropyridine calcium channel blocker. The solubilized binding sites were sensitive to pronase, alpha-chymotrypsin and phospholipases A and C indicating their protein nature as well as their lipid requirement. Chelation of endogeneous divalent cations by EDTA, EGTA or CDTA inhibits high-affinity [3H]-nimodipine binding, demonstrating that divalent cations are required for high affinity [3H]-nimodipine binding. Detergent-solubilized binding sites are adsorbed by several sepharose-immobilized lectins, including concanavalin A, wheat germ agglutinin and lentil-lectin but not by helix pomatia lectin. Preparative chromatography on concanavalin A sepharose was performed and the adsorbed [3H]-nimodipine binding sites were selectively eluted by alpha-methylmannoside; NaCl (1 mol/l) being completely ineffective as elutant. The purification factors by this method were 17-40-fold. The binding sites could be also purified (up to 10-fold) by sucrose density centrifugation. The S20, w value of the drug receptors is 12.9 s. It is concluded that the 1,4-dihydropyridine binding sites of the putative calcium channel are intimately associated with carbohydrate containing structures. Since the detergent-solubilized material shows allosteric regulation of 1,4-dihydropyridine binding, interaction with chemically different classes of calcium channel blockers, metalloprotein nature and a S20, w value which is indicative of structure large enough to span the membrane, we conclude that we have solubilized and partially purified the putative calcium channel.