Vasorelaxant mechanism of KRN2391 and nicorandil in porcine coronary arteries of different sizes.

1. The relaxant mechanisms of action of KRN2391, a novel vasodilator, and nicorandil on epimyocardial coronary artery (2.5- 3.0 mm outer diameter) and mid-myocardial coronary artery (0.8-1.0 mm outer diameter) were investigated in porcine isolated coronary arteries. In addition, the vasorelaxant responses of KRN2391 and nicorandil were compared with those of nitroglycerin and cromakalim, a K+ channel opener, in epi- and mid-myocardial coronary arteries. 2. Nitroglycerin showed a more potent relaxant effect on epi-myocardial coronary arteries than on mid-myocardial coronary arteries, whereas cromakalim produced greater relaxation responses in mid-myocardial coronary arteries. There was no difference between epi- and mid-myocardial coronary arteries in terms of the relaxant effect of KRN2391 and nicorandil. 3. Relaxation induced by KRN2391 in epi- and mid-myocardial coronary arteries was inhibited by oxyhaemoglobin, a pharmacological antagonist of nitrovasodilators, and glibenclamide, a pharmacological antagonist of K+ channel opening drugs. However, the inhibitory effect of glibenclamide on KRN2391-induced relaxation was greater in mid-myocardial coronary artery than in epi-myocardial coronary artery. 4. Relaxation induced by nicorandil was inhibited by oxyhaemoglobin alone in epi-myocardial coronary arteries and by both oxyhaemoglobin and glibenclamide in mid-myocardial coronary arteries. 5. In epi- and mid-myocardial coronary arteries, relaxation induced by cromakalim was inhibited by glibenclamide but not by oxyhaemoglobin, whereas relaxation induced by nitroglycerin was inhibited by oxyhaemoglobin but not by glibenclamide.(ABSTRACT TRUNCATED AT 250 WORDS)     

Vasospasmolytic effect of KRN2391 on 3,4-diaminopyridine-induced rhythmic contraction of porcine coronary artery

1. In the present study, we examined the vasospasmolytic effect of KRN2391 on rhythmic contractions of porcine coronary artery caused by 3,4-diaminopyridine (3,4-DAP) compared with cromakalim and nitroglycerin.2. KRN2391 at 10⁻⁷ showed a tendency to prolong the cycle length and at 10⁻⁶ M completely eliminated rhythmic contractions in all preparations. The elimination by 10⁻⁶ M KRN2391 was antagonized by either oxyhemoglobin (10⁻⁵ M) or glibenclamide (3 × 10⁻⁶) although not completely.3. Cromakalim at 10⁻⁵ M and nitroglycerin at 10⁻⁷ M completely eliminated 3,4-DAP-induced rythmic contractions in all preparations. The elimination by cromakalim and nitroglycerin was completely antagonized by glibenclamide and oxyhemoglobin, respectively.4. The present study suggests that the vasospasmolytic effect of KRN2391 on 3,4-DAP-induced rhythmic contractions is based on its nitrate action and K channel opening action.     

Vasodilator effects of KRN2391, levcromakalim and 3-morpholino-sydnonimin in human pial and omental arteries

The vasodilator action of KRN2391 (10 nM-10 microM), a combined ATP-sensitive potassium channel (KATP) opener and organic nitrate, was investigated in human pial and omental arteries. Previous animal studies have suggested that opening of KATP and activation of guanylate cyclase may contribute to varying extents to the vasodilator action of KRN2391, depending on the origin and size of the vascular preparation. Vasodilator responses were studied in isolated vascular segments (diameter 0.4-0.8 mm) pre-contracted with endothelin-1 in the presence or absence of glibenclamide (inhibitor of KATP), LY83583 (inhibitor of guanylate cyclase), zaprinast (inhibitor of cyclic GMP phosphodiesterase V) and NG-nitro-L-arginine (inhibitor of nitric oxide synthase). KRN2391 induced concentration-dependent vasodilator responses of similar potency in arteries from the two vascular regions. While glibenclamide (1 microM) had no effect in omental arteries, this compound produced a tenfold rightwards shift of the concentration-response curve for KRN2391 in pial arteries without affecting the maximal response (Emax). LY83583 (10 microM), zaprinast (10 microM) and NG-nitro-L-arginine (0.1 mM) all failed to affect the vasodilator responses to KRN2391 significantly in either artery. However, in ring segments of rat aorta LY83583 displaced the concentration-response curve for the nitric oxide donor 3-morpholino-sydnonimin (10 nM-0.1 mM) to the right, while zaprinast produced a leftwards shift. The prototype KATP opener levcromakalim (0.01-10 microM) elicited a larger relaxation in pial (Emax 80+/-6%) than in omental (Emax 47+/-13%) arteries, whereas 3-morpholino-sydnonimin produced a smaller relaxation in pial (Emax 50+/-18%) than in omental (Emax 90+/-4%) arteries. These results suggest that the vasodilator response to KRN2391 is mediated by KATP in human cerebral arteries, but dependent on neither KATP nor guanylate cyclase in human omental arteries. The results with levcromakalim and 3-morpholino-sydnonimin indicate that opening of KATP may be a more effective mechanism of vasodilatation in pial than in omental arteries from man, whereas the reverse appears to be true for guanylate cyclase activation.     

Pharmacological analysis of the inhibitory effects of KRN2391 on endothelin-1-induced contraction in isolated large coronary artery of the pig

1. The relaxant effect of KRN2391, N-cyano-N′-(2-nitroxyethyl)-3-pyridine-carboximidamide-monomethanesulfonate (with both K⁺ channel opener and nitrate actions), nifedipine (Ca²⁺ channel blocker), nitroglycerin (nitrate) and cromakalim (K⁺ channel opener) were investigated in isolated porcine large coronary arteries contracted by endothelin-1. These drugs inhibited endothelin-1-induced contraction in a concentration-dependent manner.2. The relaxation induced by KRN2391 was nearly complete at their maximum effects, but nifedipine and cromakalim could not produce complete relaxation.3. The concentration-relaxation curves for KRN2391 underwent a rightward shift in the presence of methylene blue or glibenclamide. The concentration ratios of KRN2391 calculated based on EC₅₀ values were 2.8 and 3.7 in the presence of methylene blue and glibenclamide, respectively.4. The concentration-relaxation curves for nitroglycerin and cromakalin underwent a rightward shift in the presence of methylene blue and glibenclamide, respectively, and the concentration ratios of nitroglycerin and cromakalim were 12.0 and 6.3.5. These relaxant effects of KRN2391 and nitroglycerin on endothelin-1-induced contraction of porcine coronary artery were greater than those of cromakalim and nifedipine. This potent relaxant action of KRN2391 on endothelin-induced contraction is thought to be based on both a nitrate action and a K⁺ channel opening action.     

Comparative analysis of vasodilating Mechanisms of Kil769, Ki3315 and KRN2391, pyridinecarboximidamide derivatives,in porcine isolated coronary artery

1. The vasodilating mechanisms of pyridinecarboximidamide derivatives which have a nitroxyl group (KRN2391), a phenyl group (Ki1769) or a hydroxyl group (Ki3315) were studied in porcine isolated coronary artery.2. KRN2391 (10⁻⁶ M) increased cyclic GMP formation but did not affect intracellular cyclic AMP level. Ki1769 (10⁻⁵ M) and Ki3315 (10⁻³ M) had no effect on intracellular cyclic GMP and cyclic AMP levels.3. Despite producing submaximal relaxation at KRN2391 (10⁻⁶ M) and nitroglycerin (10⁻⁶ M), the increase in cyclic GMP caused by KRN2391 was lower than that caused by nitroglycerin.4. Methylene blue (10⁻⁵ M) inhibited KRN2391- and nitroglycerin-induced relaxations but did not affect Ki1769- and Ki3315-induced relaxation.5. Glibenclamide (10⁻⁶ M) inhibited KRN2391-, Ki1769- and Ki3315-induced relaxation but did not affect nitroglycerin-induced relaxation.6. These results suggest that the nitroxyl group of KRN239 contributes to its nitrate action and the pyridinecarboximidamide moiety plays an important role of developing a K channel opening action.     

Antihypertensive and vasorelaxant effects on KRN2391 in spontaneously hypertensive rats

• 1.1. In conscious spontaneously hypertensive rats (SHR), the oral administration of KRN2391 (0.1–3.0 mg/kg) produced a dose-dependent decrease in blood pressure. The antihypertensive effect of KRN2391 was about 2 and 20 times more potent than those of pinacidil and nifedipine, respectively, but about 2 times less potent than that of cromakalim.
• 2.2. During oral administration of KRN2391 (0.5 and 1.0 mg/kg) once daily for 5 weeks, its antihypertensive effect did not diminish in conscious SHR.
• 3.3. In anaesthetized SHR, KRN2391 (3–100 μg/kg, i.v.) produced a decrease in blood pressure in a dose-dependent manner. Its antihypertensive effect was antagonized by glibenclamide (20 mg/kg, i.v.).
• 4.4. In isolated aorta obtained from SHR, KRN2391 (0.01–100 μM) produced a concentration-dependent relaxation. Its concentration-relaxation curve was shifted to the right by glibenclamide (1 μM) and methylene blue (3 μM).
• 5.5. These results indicate that the antihypertensive effect of KRN2391 in SHR is due to its direct action on vascular smooth muscle based on a K⁺ channel opening action and a nitrate action. In addition, KRN2391 is absorbed from the gastrointestinal tract into blood and does not induce tolerance despite possessing some nitrate action.

     

Comparison of KRN2391 with nicorandil and nifedipine on canine coronary blood flow: antagonism by glibenclamide.

The mode of action of KRN2391 [N-cyano-N'-(2-nitroxyethyl)-3- pyridinecarboximidamide monomethanesulfonate] in coronary circulation was examined in anesthetized dogs in comparison with those of nicorandil and nifedipine. Administration of KRN2391 (10 micrograms/kg i.v.), nicorandil (300 micrograms/kg i.v.), and nifedipine (3 micrograms/kg i.v.) caused an increase in coronary blood flow (CBF) and decreases in mean blood pressure (MBP) and in coronary vascular resistance (CVR). Heart rate (HR) was slightly and simultaneously increased by the drugs. Glibenclamide (5 mg/kg i.v.) blocked the changes of these parameters caused by KRN2391 and nicorandil, but not those caused by nifedipine. The present study suggests that the mechanism of action through ATP-sensitive K channels which are blocked by glibenclamide may contribute, at least in part, to the effects of KRN2391 and nicorandil on CBF and blood pressure (BP).     

Pharmacological analysis of the effect of KRN2391 on coronary vasculature in perfused rat heart

• 1.1. The antagonism by glibenclamide of the effects of KRN2391, cromakalim and nifedipine on coronary flow (CF) were compared in isolated perfused rat hearts.
• 2.2. KRN2391, cromakalim and nifedipine increased CF in a concentration-dependent manner. In the presence of glibenclamide, the concentration-effect curves on CF for KRN2391 and cromakalim shifted to the right but that for nifedipine did not change.
• 3.3. The EC₅₀ values increased about 6.6-fold for KRN2391 and 19.0-fold for cromakalin in the presence of glibenclamide.
• 4.4. These results suggest that the increase in CF induced by KRN2391 is due to both glibenclamide-sensitive and insensitive mechanisms.

     

Relaxant effect of trans-resveratrol on isolated porcine coronary arteries

Recent studies provided evidence that trans-resveratrol (3,4',5-trihydroxystilbene, found in high concentrations in some red wines, may possibly decrease the risk of coronary heart disease mortality. The aim of this study, performed with large epicardial porcine coronary arteries (PCA) strips, was to investigate the relaxant effect of trans-resveratrol on these main conductance vessels, which have been described to be pathologically prone for vasospastic contractions. The data show that the tonic component of the biphasic contractions induced by histamine, as well as the contractions induced by F- ions (10 mmol/l), which activate G proteins downstream of the receptors, could dose-dependently be inhibited by trans-resveratrol (0.1-100 mumol/l). The EC50 values of the dose-response curves established for the inhibition of the sustained component of histamine-induced contractions were very similar to those obtained for the relaxations of fluoride-induced contractions: 0.45 +/- 0.08 and 0.29 +/- 0.05 mumol/l, resp. (n = 6). Ouabain (10 mumol/l)-induced contractions and rhythmic contractions elicited by tetraethylammonium (12 mmol/l) were also strongly inhibited by trans-resveratrol (20 mumol/l). It may be inferred from the results obtained in this study, that the relaxation of the coronary conductance vessels induced by trans-resveratrol is possibly based on a nongenomic interaction with steroid-like receptors located on the cell membrane.     

Cardiovascular effects of KRN2391, nitroglycerin and cromakalim in dihydroergotamine-treated pithed rats

1. The effects of KRN2391 on the cardiovascular system were compared with those of nitroglycerin and cromakalim in pithed rats treated with dihydroergotamine (DHE) in order to examine the effects of these drugs on venous blood vessels.2. DHE (100 μg/kg, i.v.) produced increases in mean blood pressure (MBP), cardiac output (CO) and central venous pressure (CVP) without changes in total peripheral vascular resistance (TPR) and heart rate (HR) based on venoconstriction. The DHE-treated pithed rats, nitroglycerin (30 μg/kg, i.v.) decreased CO and CVP whereas cromakalim (30 μg/kg, i.v.) produced a slight increase in CO followed by a decrease and did not affect CVP. KRN2391 (30 μg/kg, i.v.) produced a decrease in CVP without affecting CO. Decreases in MBP and TPR were induced by all drugs.3. These results suggest that nitroglycerin acts predominantly as a venodilator and KRN2391 and cromakalim showed a venodilating action in addition to an arterial dilating action in DHE treated pithed rats. However, the venodilating action of KRN2391 in this condition is more potent than that of cromakalim.     

Effect of KRN2391 on canine ventricular arrhythmia models

• 1.1. KRN2391 (3–30 μg/kg, i.v.) produced a decrease in mean blood pressure (MBP) with concomitant increase in heart rate (HR) and change in electrocardiogram (ECG) such as the shortening of PP and PQ intervals and the prolongation of QTc and these changes in HR and ECG were attenuated by pretreatment with propranolol (1 mg/kg) in normal dogs.
• 2.2. KRN2391 at 30 μg/kg induces neither suppression nor aggravation of ventricular arrhythmias caused by adrenaline and digitalis.
• 3.3. In two-stage coronary ligation-induced arrhythmia, KRN2391 inhibited arrhythmia at 48 hr.
• 4.4. These results suggest that KRN2391 may be effective on arrhythmia related to ischemia. In addition, it is considered that arrhythmia is not induced even by a high dose of KRN2391 in the normal condition.

     

Vasodilatory effects of lidocaine on epicardial porcine coronary arteries

To investigate the mechanism of lidocaine's effect to cause vasorelaxation, swine epicardial mid-right coronary arterial rings were placed under constant (5 g) tension in a muscle bath, precontracted with 35 mmol/l KCl and exposed to increasing concentrations of lidocaine (3-2,000 micrograms/ml). At a concentration of 10 micrograms/ml, mild vasoconstriction occurred, increasing tension 1.9 +/- 0.1% above baseline. Vasodilation began to occur at 30 micrograms/ml and was maximal at 2,000 micrograms/ml, reducing tension 97.5 +/- 0.2% below baseline. Vasodilation was not altered significantly by removal of endothelium or by pretreatment with propranolol or indometacin.     

Cardioprotective effects of KRN2391 and nicorandil on ischemic dysfunction in perfused rat heart.

The cardioprotective effect of KRN2391 (N-cyano-N-(2-nitroxymethyl)-3- pyridinecarboximidamide methanesulfonate), a novel vasodilator, was studied in the isolated perfused rat heart and compared with that of nicorandil. The isolated buffer-perfused rat heart was subjected to 25 min ischemia followed by 30 min reperfusion. The heart was pretreated with 0.1-10 microM KRN2391, 10-1000 microM nicorandil or vehicle. Before ischemia, KRN2391 (1-10 microM) and nicorandil (10-1000 microM) increased coronary flow, but did not modify the cardiac mechanical function. KRN2391 (1-10 microM) and nicorandil at high doses (300-1000 microM) resulted in significant improvements of cardiac functions and coronary flow during reperfusion and significantly reduced the release of cytosolic enzymes. The protective effects of 3 microM KRN2391 and 300 microM nicorandil were completely reversed by 3 microM glibenclamide, a blocker of ATP-sensitive potassium channels. Thus, KRN2391 and nicorandil at high doses have a direct cardioprotective effect, which may be related to activation of ATP-sensitive potassium channels.     

Normothermic and hypothermic models for studying the deleterious effects of hypoxia-reoxygenation on EDHF-mediated relaxation in isolated porcine coronary arteries

IntroductionThe vasomotor response of the coronary artery is altered by hypoxia–reoxygenation (H–R) induced damage. The aim of our study was to compare and evaluate normothermic and hypothermic models which are suitable for future drug studies of vasoprotective action against H–R injury.MethodsPorcine coronary arterial rings were isolated and placed in Krebs–Henseleit (K–H) solution. Rings were exposed to normoxic conditions (control group) and two different H–R conditions: the first induced by a 95% N₂–5% CO₂ gas mixture (40- and 60-min hypoxia) in a normothermic protocol, and the second induced by hypothermic (4 °C) hypoxia–reoxygenation in an air-tight beaker filled with K–H solution (24- and 48-hours hypoxia). Reoxygenation was applied by introducing K–H solution aerated with a 95% O₂–5% CO₂ mixture under normothermic (37 °C) conditions. To test the EDHF-mediated relaxation by substance P, rings were first incubated in L-NNA, nitric oxide synthase inhibitor, and indomethacin, cyclooxygenase inhibitor, and then pre-contracted with thromboxane analogue U-46619. Analysis of the maximum relaxation of the arterial rings was performed by one-way ANOVA, followed by Bonferroni's post-test.ResultsDistal segments of the coronary artery responded faster to contraction induced by U-46619 and were relaxed by substance P to a greater extent than proximal segments. Maximal relaxations of arterial rings induced by a 10 nM solution of substance P were significantly reduced (p < 0.001) from the values for normoxic rings (81.0 ± 1.0%, n = 30) after 40-min H–R (50.5 ± 5.3%, n = 30), 60-min H–R (32.1 ± 3.5%, n = 30), 24-hours hypothermic H–R (56.0 ± 2.3%, n = 30) and after 48-hours hypothermic H–R (38.5 ± 5.1%, n = 30).ConclusionsThe model employing 40-min normothermic H–R is as effective as 24-hours hypothermic H–R, and 60-min normothermic H–R as 48-hours hypothermic H–R for studying the deleterious effects of H–R on EDHF-mediated relaxation.     

Contribution of cyclic GMP formation to KRN2391-induced relaxation in coronary artery of the pig.

1. In the present study, we investigated the relationship between relaxation and guanosine 3':5'-cyclic monophosphate (cyclic GMP) formation induced by KRN2391, compared with those induced by nicorandil and nitroglycerin, in the coronary artery of the pig. 2. KRN2391 (10(-8)-3 X 10(-5) M), nicorandil (10(-8)-3 X 10(-4) M) and nitroglycerin (10(-9)-10(-5) M) antagonized the contraction caused by 25 mM KCl in a concentration-dependent manner. 3. The concentration-relaxation curves for KRN2391, nicorandil and nitroglycerin shifted rightward in the presence of methylene blue (10(-5) M). 4. KRN2391 (10(-6) M), nicorandil (10(-4) M) and nitroglycerin (10(-6) M) induced an increased in cyclic GMP. 5. The magnitude of the shift of the concentration-relaxation curve caused by methylene blue and the increase in cyclic GMP with KRN2391 were lower than those with nicorandil and nitroglycerin. 6. The adenosine 3':5'-cyclic monophosphate (cyclic AMP) level was not increased by KRN2391 even at a concentration that produced full relaxation. 7. The present results suggest that KRN2391-induced relaxation in the coronary artery of the pig is partly due to the increase in cyclic GMP formation through the stimulation of guanylate cyclase.     

Effects of KRN2391 on ionic currents in rabbit femoral arterial myocytes

The effects of KRN2391, an ATP-sensitive K+ channel opener (KCO) which also acts as a nitrate, on ionic membrane currents in rabbit femoral arterial myocytes were examined. Under whole-cell clamp conditions where cells were superfused with physiological salts solution containing 5.9 mM K+, KRN2391 elicited an outward current at a holding potential of -30 mV. KRN2391-induced current had a reversal potential of -78 mV and was abolished by glibenclamide (glib). KRN2391 was approximately 25 times more potent than nicorandil to activate an ATP-sensitive K+ current (I:(KATP)). On the other hand, 10 microM KRN2391 did not affect either voltage-dependent Ca(2+) or delayed rectifier K+ channel currents. In the inside-out patch configuration, KRN2391 activated 47 pS K+ channels in the presence of nucleotide diphosphates (NDPs) under the symmetrical 140 mM K+ conditions. Glib and intracellular ATP reversibly inhibited the activity of the 47 pS K+ channels. The 47 pS K+ channels activated by KRN2391 are similar in their conductance and other properties to NDP-sensitive K+ channels (K(NDP) channels) described in other smooth muscles and the cloned channels. KRN2391 is a potent activator of the 47 pS K+ channels and the activation can contribute to the KRN2391-induced vasodilation in arterial muscles.     

Comparison of the inhibitory effects of nifedipine and nimodipine on mechanical responses of isolated rat coronary small arteries

The effect of nifedipine and nimodipine was studied on the spontaneous myogenic tone and on the serotonin-, potassium-, and calcium-induced contractile responses in coronary resistance arteries from 2-year-old male Wistar rats. Both drugs depressed completely and concentration-dependently the spontaneous myogenic tone. At the highest concentration, nifedipine and nimodipine reduced the serotonin and potassium responses to an average of 19% and 6% of control response. The drugs had an equipotent inhibitory effect on the serotonin response. Nimodipine, however, was more potent than nifedipine, inhibiting the potassium response. In vessels maximally activated by serotonin and potassium, both drugs concentration-dependently inhibited the calcium concentration-response curves. Nifedipine and nimodipine had an equieffective and equipotent inhibitory effect on the calcium responses in serotonin-activated vessels. Nimodipine was more potent than nifedipine, inhibiting the calcium response of potassium-activated vessels. In the highest concentration, nimodipine was also more effective than nifedipine (p less than 0.01) in inhibiting the calcium response. In conclusion, both nifedipine and nimodipine are potent and effective vasodilators, having similar actions on the spontaneous myogenic tone and serotonin-induced responses in the rat coronary resistance arteries, but nimodipine is more potent and more effective than nifedipine in inhibiting potassium-induced responses.     

Pharmacological analysis of the vasodepressor effect of KRN2391 in pithed rats

• 1.1. The antagonism by glibenclamide of the vasodepressor effects of KRN2391, cromakalim and nitroglycerin was compared in pithed rats with blood pressure supported by an infusion of phenylephrine.
• 2.2. Cumulative administration of KRN2391 (3–300 μg/kg, i.v.), cromakalim (3–300 μg/kg, i.v.) and nitroglycerin (1–300 μg/kg, i.v.) produced dose-dependent decreases in diastolic blood pressure.
• 3.3. In rats given glibenclamide (20 mg/kg, i.v.), the dose-vasodepressor curves for KRN2391 and cromakalim were shifted to the right. However, glibenclamide had no effect on the vasodepressor effect of nitroglycerin.
• 4.4. The ED_{50 mmHg} values increased about 5.9 fold for KRN2391 and 9.5 fold for cromakalim in glibenclamide-treated rats.
• 5.5. These results suggest that the vasodepressor effect of KRN2391 is due to both glibenclamide-sensitive and insensitive mechanisms. This glibenclamide-insensitive effect of KRN2391 is thought to reflect its nitrate action.

     

Comparative effects of the isomers of bepridil on isolated coronary and aortic arteries

The vasodilator actions of racemic bepridil were compared with those of its laevo-(l) and dextro-(d) rotatory isomers in isolated rabbit aorta and pig coronary artery. The actions of bepridil (B), (l) B and (d) B were further compared with those of drugs known to act either by blockade of calcium entry or to inhibit calmodulin in pig coronary artery. (l) B and (d) B were equipotent in relaxing tonic contractions induced by phenylephrine in rabbit aorta but (d) B was approximately twice as potent as (l) B in relaxing tonic contractions induced by potassium (K+). Both (d) B and (l) B relaxed K+-induced contractions in coronary artery and, in higher concentrations, inhibited and N-(6-aminohexyl)-5-chloro-1-naphthalene-sulfonamide (W7) were equipotent against both types of contraction whilst nifedipine and verapamil failed to reduce histamine-induced contractions. Both isomers of bepridil (like W7) shifted concentration-response curves to histamine in non-depolarized coronary artery in a noncompetitive manner. No potency differences were found between (l) B and (d) B in this tissues. It is concluded that intracellular actions, possibly calmodulin inhibition, play a substantial role in the vasodilator action of bepridil, a conclusion supported by the relative lack of stereospecificity shown by the bepridil isomers.