(Circulation. 1995;91:262-264.)
© 1995 American Heart Association, Inc.
Articles |
ß-Adrenergic Blocking Property of dl-Sotalol Maintains Class III Efficacy in Guinea Pig Ventricular Muscle After Isoproterenol
Presented in part at the Annual Meeting of the North American Society of Pacing and Electrophysiology, Nashville, Tenn, May 11-14, 1994.
From the Division of Cardiology, Department of Medicine (W.J.G., J.H.M.) and the Department of Obstetrics and Gynecology (K.J.G., J.G.M.), Oregon Health Sciences University, Portland.
Correspondence to James G. Maylie, PhD, Oregon Health Sciences University M/S L468, 3181 SW Sam Jackson Park Rd, Portland, OR 97201-3098.
 |
Abstract |
|---|
 Top Abstract IntroductionMethods Results Discussion References |
|---|
Background Catecholamines antagonize the efficacy of several class III antiarrhythmic agents. To determine the role of the intrinsic ß-adrenergic blocking property of dl-sotalol in maintaining class III efficacy during a high-catecholamine state, we compared the electrophysiological properties of dl-sotalol with those of d-sotalol, which is devoid of significant ß-adrenergic blocking effect, before and after isoproterenol infusion.
Methods and Results Action potential duration at 90% repolarization (APD90) was prolonged in isolated guinea pig papillary muscles perfused with d-sotalol and dl-sotalol 10-4 mol/L over stimulation cycle lengths from 200 to 2000 ms. The increases in APD90 for d-sotalol and dl-sotalol over control were 10.9±2.5 to 23.7±4.8 ms and 27.9±4.0 to 39.0±5.6 ms, respectively. APD90 shortened to less than control in papillary muscles treated with d-sotalol but not dl-sotalol on addition of isoproterenol 10-6 mol/L: -31.2±3.5 to -18.3±4.8 ms and 10.5±3.6 to 33.3±7.8 ms, respectively, P<.003. Single guinea pig ventricular myocytes were studied by the whole-cell patch clamp method. Time-dependent (Iout) and total (Itot) outward current in response to a 300-ms pulse to 20 mV and tail current (Itail) to -35 mV were measured after Ca2+ channel block and Na+ channel inactivation. Iout, Itail, and Itot were reduced in myocytes perfused with d-sotalol and dl-sotalol 10-4 mol/L: Iout, -36.1±4.1%, -40.5±3.3%; Itail, -59.3±4.6%, -62.2±11.1%; Itot, -27.3±4.3%, -50.0±11.8%. Iout and Itot increased to a greater degree in myocytes treated with d-sotalol than dl-sotalol on addition of isoproterenol 10-6 mol/L: Iout, 100.3±20.6%, 11.3±7.6%, P=.002; Itot, 86.8±39.2%, -41.1±20.9%, P=.01. Itail tended to increase more in myocytes treated with d-sotalol than dl-sotalol on addition of isoproterenol, but the difference was not significant (-9.1±13.5%, -28.0±9.0%).
Conclusions The ß-adrenergic blocking property of dl-sotalol maintains APD prolongation and repolarizing outward current block during isoproterenol infusion in guinea pig ventricular muscle. Extrapolation of these data to a clinical setting may explain the efficacy of dl-sotalol in diminishing ventricular arrhythmia recurrence.
Key Words: antiarrhythmia agents • potassium • catecholamines
|
Introduction |
|---|
|
TopAbstractIntroductionMethods Results Discussion References |
|---|
The drug dl-sotalol is effective therapy for ventricular tachyarrhythmias.1 As a class III antiarrhythmic agent, the mechanism of action of dl-sotalol is to block repolarizing outward current, prolonging cardiac action potential duration (APD) and refractory period and thereby reducing the likelihood of reentrant arrhythmias.2 3 4 5 6 dl-Sotalol blocks a rapid component of the delayed rectifier K+ current, IKr7 In addition, dl-sotalol has significant ß-adrenergic blocking property. In contrast, d-sotalol, an equipotent IKr blocker, has minimal ß-adrenergic blocking property.8 A slow component of the delayed rectifier K+ current, IKs, and a Cl- current (ICl) are upregulated by the ß-adrenergic agonist isoproterenol.9 10 Upregulation of these outward currents can counteract the APD-prolonging effects of IKr blocking agents. Pretreatment with ß-adrenergic blockers maintains the APD- and refractory period–prolonging effects of IKr blocking agents exposed to isoproterenol.9 This study examines the role of the intrinsic ß-adrenergic blocking property of dl-sotalol in maintaining class III effectiveness after isoproterenol.
|
Methods |
|---|
|
TopAbstractIntroductionMethods Results Discussion References |
|---|
Twenty-nine guinea pigs (200 to 300 g, male) were used for this study. Animal preparation and handling were performed according to the guiding principles of the American Society of Physiology and under a protocol approved by the Animal Care Committee at Oregon Health Sciences University. Guinea pigs were anesthetized with halothane inhalation, and the hearts were removed.
Papillary Muscle Preparation
A right ventricular papillary
muscle was dissected free, mounted
in a muscle bath, and superfused with 1.8 mmol/L
Ca2+-Tyrode's solution (in mmol/L: NaCl 140, KCl
5.4, MgCl2 1, glucose 10, HEPES 10, adjusted with NaOH to
pH 7.35) gassed with 100% O2 and maintained at a
temperature of 37.0±0.2°C.
Muscle strips were attached to a
force transducer, and resting length
was adjusted to produce maximum isometric force. The papillary muscle
was stimulated with a pulse width of 1 ms and voltage of twice
threshold. Intracellular APs were recorded on an electronic strip
recorder (MacLab, AD Instruments) with 20 to 30 M
3 mol/L KCl-filled
microelectrodes (A-M Systems, Inc). APs were recorded at stimulation
cycle lengths from 200 to 2000 ms before and 30 minutes after the
addition of d- or dl-sotalol 10-4
mol/L (Bristol-Myers Squibb Pharmaceuticals) and again 30 minutes after
the addition of isoproterenol 10-6 mol/L (Elkins-Sinn). A
dose of 10-4 mol/L sotalol was used to maximally block
IKr without significant effect on other
currents.4
Ventricular Myocyte Isolation
Ventricular myocytes were
isolated as previously
described.11 Hearts were perfused retrogradely with 100%
O2/Ca2+-free Tyrode's solution
for 5 minutes. The solution was changed to one containing collagenase
(type II, 275 U/mL; Worthington Biochemical Corp) and protease (type
XIV, 0.8 U/mL; Sigma Chemical Co) for 7 minutes. The heart was
subsequently perfused with 0.1 mmol/L Ca2+-Tyrode's
solution for 10 minutes. Ventricles were dissected, minced, and placed
in 0.1 mmol/L Ca2+-Tyrode's solution. Cells were
studied within 8 hours of dissociation.
Whole-Cell Current Recording
Whole-cell currents were
recorded from calcium-tolerant myocytes
with patch electrodes (A-M Systems, Inc) with resistances of 2.5 to 4
M when filled with (in mmol/L): potassium glutamate 80, KCl 40, NaCl
10, MgCl2 1, MgATP 5, CaCl2 1, potassium
creatine phosphate 0.5, EGTA 10, and HEPES 10, adjusted with KOH to pH
7.1. Cells were voltage-clamped with an Axopatch 1B amplifier (Axon
Instruments). Series resistance was 50% to 70% compensated, and the
currents were filtered at 1 kHz before digitization and storage on an
LSI 1173 computer (Digital Equipment Corp).
Cells were perfused at a constant rate of 1.5 mL/min with 1.8 mmol/L Ca2+-Tyrode's solution held at 29.0±0.2°C. Ca2+ channels were blocked with 1 to 2x10-4 mol/L Cd2+, and Na+ channels were inactivated with a 1000-ms prepulse to -40 mV. Myocytes were held at their resting membrane potential of -72 to -75 mV and depolarized at 1-minute intervals to 20 mV for 300 ms, a time frame and voltage representative of the intrinsic action potential. Time-dependent outward current (Iout) was measured as the difference between final and initial current. Total outward current (Itot) equals the final current and reflects the sum of instantaneous and time-dependent outward current. Tail current (Itail) was measured at -35 mV, near the Cl- equilibrium potential. After control determination, either d-sotalol or dl-sotalol 10-4 mol/L was added to the perfusate, and data were collected for 10 minutes. Isoproterenol 10-6 mol/L was then added to the drug perfusate, and data were collected again for 10 minutes. In all experiments, steady state was achieved during the 10-minute collection times.
Data Analysis
The time difference in APD to 90%
repolarization
(APD90) between control and each drug state was calculated
for each pacing cycle length. Current measurements were expressed as
percent change from control (100%x[drug
state-control]/control),
which corrected for differences in cell size. Effects on APD in
papillary muscles and currents in myocytes treated with d-
or dl-sotalol were compared by a two-tailed
independent-samples t test. This comparison was made before
and after isoproterenol infusion. Values of P<.05 were
taken as statistically significant.
|
Results |
|---|
|
TopAbstractIntroductionMethods Results Discussion References |
|---|
Results are summarized in the Table
.
|
Papillary Muscles
APD90 was rate-dependent,
increasing by 114.6±5.0 to
156.5±8.4 ms from the pacing cycle lengths of 200 to 2000 ms,
respectively. Results are shown in Fig 1. Both
d- and dl-sotalol 10-4 mol/L
prolonged the APD without effect on the maximal rate of rise of the AP.
There was a trend for dl-sotalol to prolong
APD90 to a greater extent than d-sotalol, but
this was not significant. APD shortened to less than control in muscle
treated with d-sotalol but not dl-sotalol on
addition of isoproterenol 10-6 mol/L.
|
Isolated Guinea Pig Ventricular Myocytes
Iout,
Itot, and
Itail in response to a 300-ms pulse to 20 mV with tail to
-35 mV averaged 50.6±5.5, 74.7±16.9, and 19.1±2.8
pA, respectively.
Results are shown in Fig 2. Iout,
Itot, and Itail were reduced in myocytes
perfused with d- and dl-sotalol. There was a
trend for dl-sotalol to reduce Itot to a greater
extent than d-sotalol, but this was not significant.
Iout increased compared with control on addition of
isoproterenol in myocytes treated with both d- and
dl-sotalol but to a significantly greater extent in those
treated with d-sotalol. Itot remained reduced
below control in myocytes treated with dl-sotalol but not
d-sotalol. Itail increased but not to control
values with addition of isoproterenol in myocytes treated with either
d-sotalol or dl-sotalol. There was a trend for
Itail to increase to a greater extent with addition of
isoproterenol in myocytes treated with d-sotalol compared
with dl-sotalol, but the difference was not
significant.
|
|
Discussion |
|---|
|
TopAbstractIntroductionMethods Results Discussion References |
|---|
dl-Sotalol but not d-sotalol maintains the class III effects of APD prolongation and diminished outward current during isoproterenol infusion in guinea pig ventricular muscle. This differential effect is secondary to the ß-adrenergic blocking property of dl-sotalol counteracting the isoproterenol-induced increase in total outward current. In myocytes treated with d-sotalol, an instantaneous current, consistent with ICl, and an increased time-dependent current, consistent with an increase in IKs, occurred with isoproterenol infusion. These isoproterenol-induced currents increased total outward current, overwhelming the effect of IKr block by d-sotalol. Catecholamines antagonize the class III efficacy of several antiarrhythmic agents known to block IKr in humans, consistent with our findings for d-sotalol in guinea pig heart.12 13 14 15 Ionic currents responsible for repolarization in human heart have not been well characterized, and therefore, the mechanism by which catecholamines antagonize class III antiarrhythmic efficacy may be different from that in guinea pig heart.
With isoproterenol challenge, Itail did not increase to control value with either d-sotalol or dl-sotalol, and the difference between drugs was not significant. This is probably due to IKr inward rectification,7 which increases the contribution of IKr to Itail compared with Iout.
d-Sotalol and other pure IKr blockers, despite potent class III effects, may be limited in their efficacy by high-catecholamine states. The clinical efficacy of dl-sotalol may be related to the combination of class III effects, which prolong APD and thereby reduce the likelihood of reentrant arrhythmias, and ß-adrenergic blocking property, which maintains class III efficacy despite high-catecholamine states.
|
Acknowledgments |
|---|
This research was supported in part by an American Heart Association, Oregon Affiliate, grant (Dr Groh) and by National Institutes of Health grant HL-48286 (Dr Maylie).
Received September 26, 1994; accepted November 25, 1994.
|
References |
|---|
|
TopAbstractIntroductionMethods Results Discussion References |
|---|
1. Mason JW. A comparison of seven antiarrhythmic drugs in patients with ventricular tachyarrhythmias. N Engl J Med. 1993;329:452-458.
2. Singh BN, Vaughan Williams EM. A third class of anti-arrhythmic action: effects on atrial and ventricular intracellular potentials, and other pharmacologic actions on cardiac muscle, of MJ 1999 and AH 3474. Br J Pharmacol. 1970;39:675-687. [Medline] [Order article via Infotrieve]
3.
Strauss HC, Bigger JT Jr, Hoffman BF. Electrophysiological
and beta-receptor blocking effects of MJ 1999 on dog and rabbit cardiac
tissue. Circ Res. 1970;26:661-678.
4.
Carmeliet E. Electrophysiologic and voltage clamp
analysis of the effects of sotalol on isolated cardiac muscle and
Purkinje fibers. J Pharmacol Exp Ther. 1985;232:817-825.
5. Echt DS, Berte LE, Clusin WT, Samuelsson RG, Harrison DC, Mason JW. Prolongation of the human cardiac monophasic action potential by sotalol. Am J Cardiol. 1982;50:1082-1086. [Medline] [Order article via Infotrieve]
6. Singh BN, Nademanee K. Control of cardiac arrhythmias by selective lengthening of repolarization: theoretic considerations and clinical observations. Am Heart J. 1985;109:421-430. [Medline] [Order article via Infotrieve]
7.
Sanguinetti MC, Jurkiewicz NK. Two components of cardiac
delayed rectifier K+ current: differential sensitivity to
block by class III antiarrhythmic agents. J Gen Physiol. 1990;96:195-215.
8. Kato R, Ikeda N, Yabek SM, Kannan R, Singh BN. Electrophysiologic effects of the levo- and dextrorotatory isomers of sotalol in isolated cardiac muscle and their in vivo pharmacokinetics. J Am Coll Cardiol. 1986;7:116-125. [Abstract]
9.
Sanguinetti MC, Jurkiewicz NK, Scott A, Siegl PK.
Isoproterenol antagonizes prolongation of refractory period by the
class III antiarrhythmic agent E-4031 in guinea pig myocytes: mechanism
of action. Circ Res. 1991;68:77-84.
10.
Harvey RD, Clark CD, Hume JR. Chloride current in mammalian
cardiac myocytes: novel mechanism for autonomic regulation of action
potential duration and resting membrane potential. J Gen
Physiol. 1990;95:1077-1102.
11.
Zygmunt AC, Maylie J. Stimulation-dependent facilitation of
the high threshold calcium current in guinea-pig ventricular myocytes.
J Physiol (Lond). 1990;428:653-671.
12. Wellens HJ, Brugada P, Roy D, Weiss J, Bar FW. Effect of isoproterenol on the anterograde refractory period of the accessory pathway in patients with the Wolff-Parkinson-White syndrome. Am J Cardiol. 1982;50:180-184. [Medline] [Order article via Infotrieve]
13. Brugada P, Facchini M, Wellens HJ. Effects of isoproterenol and amiodarone and the role of exercise in initiation of circus movement tachycardia in the accessory atrioventricular pathway. Am J Cardiol. 1986;57:146-149. [Medline] [Order article via Infotrieve]
14.
Brembilla-Perrot B, Admant PH, Le Helloco A, Pernot C. Loss of
efficacy of flecainide in the Wolff-Parkinson-White syndrome after
isoproterenol administration. Eur Heart J. 1985;6:1074-1078.
15.
Keren A, Tzivoni D, Gavish D, Levi J, Gottlieb S, Benhorin J,
Stern S. Etiology, warning signs and therapy of torsade de pointes: a
study of 10 patients. Circulation. 1981;64:1167-1174.
This article has been cited by other articles:
 |
|
 |
|
  C.-E. Chiang, H.-N. Luk, and T.-M. Wang Swelling-activated chloride current is activated in guinea pig cardiomyocytes from endotoxic shock Cardiovasc Res, April 1, 2004; 62(1): 96 - 104. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C.-E. Chiang, H.-N. Luk, T.-M. Wang, and P. Y.-A. Ding Prolongation of cardiac repolarization by arsenic trioxide Blood, August 28, 2002; 100(6): 2249 - 2252. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Sen, G. Cui, Y. Sakaguchi, and B. N. Singh Electrophysiological Effects of MS-551, a New Class III Agent: Comparison with dl-Sotalol in Dogs J. Pharmacol. Exp. Ther., May 1, 1998; 285(2): 687 - 694. [Abstract] [Full Text]
|
|
|
|
 |
|
 S. H. Hohnloser, T. Meinertz, P. Stubbs, H. J.G.M. Crijns, J.-J. Blanc, P. Rizzon, and B. Cheuvart Efficacy and Safety of d-Sotalol, a Pure Class III Antiarrhythmic Compound, in Patients With Symptomatic Complex Ventricular Ectopy : Results of a Multicenter, Randomized, Double-blind, Placebo-Controlled Dose-Finding Study Circulation, September 15, 1995; 92(6): 1517 - 1525. [Abstract] [Full Text]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||