Rate-dependent Drug Effects Research Articles

Mechanisms Underlying Rate-Dependent Effects of State-Specific Binding of Sodium Channel Blockers in Cardiac Tissue: Insights from Idealized Models

Increased propensity for arrhythmias is a well-documented but poorly understood side-effect of drugs. Many arrhythmogenic drugs bind to and block ion-channels in a conformational state-specific manner, making it difficult to discern the fundamental mechanisms underlying pro-arrhythmic drug effects. We have developed mathematical models to examine the effects of state-specific Na+ channel blockers with various binding mechanisms on the rate-dependence of peak upstroke velocity, conduction velocity, and the vulnerable window of cardiac tissue. Specifically, we compared the effects of drugs that bind via the guarded receptor and gate immobilization models, which are idealized models consisting of motifs commonly found in modulated receptor models of drug binding. For both the guarded receptor and gate immobilization models, we considered drugs that bind to either inactivated or non-inactivated Na-channels. Our results suggest that drugs that bind to inactivated Na+ channels give rise to greater rate-dependent effects on peak upstroke velocity and conduction velocity than drugs that bind to non-inactivated Na+ channels. Additionally, we find drugs that bind to non-inactivated Na+ channels via the gate immobilization motif induce a reversal of the rate-dependence of peak upstroke velocity. By exploiting the idealized nature of guarded receptor and gate immobilization models and utilizing singular perturbation analysis, we provide an explanation of the fundamental mechanisms of rate-dependent drug effects. Finally, we show that a gate immobilization model with inactivated state binding captures the rate-dependent effects of lidocaine, demonstrating that idealized models can be useful models to study mechanisms of action of real drugs.

Frequency-Dependent Multi-Well Cardiotoxicity Screening Enabled by Optogenetic Stimulation.

Side effects on cardiac ion channels causing lethal arrhythmias are one major reason for drug withdrawals from the market. Field potential (FP) recording from cardiomyocytes, is a well-suited tool to assess such cardiotoxic effects of drug candidates in preclinical drug development, but it is currently limited to the spontaneous beating of the cardiomyocytes and manual analysis. Herein, we present a novel optogenetic cardiotoxicity screening system suited for the parallel automated frequency-dependent analysis of drug effects on FP recorded from human-induced pluripotent stem cell-derived cardiomyocytes. For the expression of the light-sensitive cation channel Channelrhodopsin-2, we optimised protocols using virus transduction or transient mRNA transfection. Optical stimulation was performed with a new light-emitting diode lid for a 96-well FP recording system. This enabled reliable pacing at physiologically relevant heart rates and robust recording of FP. Thereby we detected rate-dependent effects of drugs on Na+, Ca2+ and K+ channel function indicated by FP prolongation, FP shortening and the slowing of the FP downstroke component, as well as generation of afterdepolarisations. Taken together, we present a scalable approach for preclinical frequency-dependent screening of drug effects on cardiac electrophysiology. Importantly, we show that the recording and analysis can be fully automated and the technology is readily available using commercial products.

Mechanism of Reverse Rate-Dependent Action of Cardioactive Agents

Class 3 antiarrhythmic agents exhibit reverse rate-dependent lengthening of the action potential duration (APD), i.e. changes in APD are greater at longer than at shorter cycle lengths. In spite of the several theories developed to explain this reverse rate-dependency, its mechanism has been clarified only recently. The aim of the present study is to elucidate the mechanisms responsible for reverse rate-dependency in mammalian ventricular myocardium. Action potentials were recorded using conventional sharp microelectrodes from human, canine, rabbit, guinea pig, and rat ventricular myocardium in a rate-dependent manner. Rate-dependent drug-effects of various origin were studied using agents known to lengthen or shorten action potentials allowing thus to determine the drug-induced changes in APD as a function of the cycle length. Both drug-induced lengthening and shortening of action potentials displayed reverse rate-dependency in human, canine, and guinea pig preparations, but not in rabbit and rat myocardium. Similar results were obtained when repolarization was modified by injection of inward or outward current pulses in isolated canine cardiomyocytes. In contrast to reverse rate-dependence, drug-induced changes in APD well correlated with baseline APD values (i.e. that measured before the superfusion of drug or injection of current) in all of the preparations studied. Since the net membrane current (I(net)), determined from the action potential waveform at the middle of the plateau, was inversely proportional to APD, and consequently to cycle length, it is concluded that that reverse rate-dependency may simply reflect the inverse relationship linking I(net) to APD. In summary, reverse rate-dependency is an intrinsic property of drug action in the hearts of species showing positive APD - cycle length relationship, including humans. This implies that development of a pure K(+) channel blocking agent without reverse rate-dependent effects is not likely to be successful.

Reverse rate-dependent changes are determined by baseline action potential duration in mammalian and human ventricular preparations

Class III antiarrhythmic agents exhibit reverse rate-dependent lengthening of the action potential duration (APD). In spite of the several theories developed so far to explain this reverse rate-dependency (RRD), its mechanism has not yet been clarified. The aim of the present work was to further elucidate the mechanisms responsible for RRD in mammalian ventricular myocardium. Action potentials were recorded using conventional sharp microelectrodes from human, canine, rabbit and guinea pig ventricular myocardium in a rate-dependent manner varying the cycle length (CL) between 0.3 and 5 s. Rate-dependent drug effects were studied using agents known to lengthen or shorten action potentials, and these drug-induced changes in APD were correlated with baseline APD values. Both drug-induced lengthening (by dofetilide, sotalol, E-4031, BaCl(2), veratrine, BAY K 8644) and shortening (by mexiletine, tetrodotoxin, lemakalim) of action potentials displayed RRD, i.e., changes in APD were greater at longer than at shorter CLs. In rabbit, where APD is a biphasic function of CL, the drug-induced APD changes were proportional to baseline APD values but not to CL. Similar results were obtained when repolarization was modified by injection of inward or outward current pulses in isolated canine cardiomyocytes. In each case the change in APD was proportional to baseline APD (i.e., that measured before the superfusion of drug or injection of current). Also, the net membrane current (I (net)), determined from the action potential waveform at the middle of the plateau, was inversely proportional to APD and consequently with to CL. The results indicate that RRD is a common characteristic of all the drugs tested regardless of the modified ion current species. Thus, drug-induced RRD can be considered as an intrinsic property of cardiac membranes based on the inverse relationship between I (net) and APD.

A method for QT correction based on beat-to-beat analysis of the QT/RR interval relationship in conscious telemetred beagle dogs

Drug-induced QT prolongation is a major clinical risk factor for arrhythmia induction, particularly torsades de pointes. QT interval is rate dependent, and many formulae exist that attempt to correct QT for changes in heart rate. Most correction factors are acknowledged to overcorrect at high heart rates, undercorrect at low heart rates, and tend to be species specific. Data collected from computerised data acquisition systems are normally reported as means over a given logging period, and so extremes of heart rate are averaged out. Therefore, the aim of this study was to develop a technique for assessing drug-induced changes in the QT/RR relationship, which is simple, suitable for small group sizes, and better able to determine rate-dependent effects of drugs. Telemetred beagle dogs (n=4) instrumented for the measurement of electrocardiogram (ECG) were monitored for four separate 20-h periods to define the control QT/RR relationship. Data were binned by RR interval, in 10 ms bins, to produce a control curve. Each dog was treated with vehicle and sotalol (4, 8, 32 mg/kg) in a crossover design to determine whether drug-induced changes in the QT/RR relationship could be detected using the data binning technique. The control QT/RR relationship was curvilinear with a steep section for RR intervals below 580 ms, and was much less steep after this point. Sotalol produced QT prolongation and bradycardia-Fridericia's correction (QTf) reduced the magnitude of this prolongation. The data analysed by the binning technique showed a larger prolongation in QT than was suggested by QTf, and an inverse frequency-dependent response. Beat-to-beat analysis and binning allows accurate determination of the QT/RR relationship and assessment of QT prolongation without recourse to mathematical modelling. It also highlights the importance of assessing QT effects in well-trained animals over a range of heart rates.

K+ channel structure-activity relationships and mechanisms of drug-induced QT prolongation.

Pharmacological intervention, often for the purpose of treating syndromes unrelated to cardiac disease, can increase the vulnerability of some patients to life-threatening rhythm disturbances. This may be due to an underlying propensity stemming from genetic defects or polymorphisms, or structural abnormalities that provide a substrate allowing for the initiation of arrhythmic triggers. A number of pharmacological agents that have proven useful in the treatment of allergic reactions, gastrointestinal disorders, and psychotic disorders, among others, have been shown to reduce repolarizing K(+) currents and prolong the QT interval on the electrocardiogram. Understanding the structural determinants of K(+) channel blockade may provide new insights into the mechanism and rate-dependent effects of drugs on cellular physiology. Drug-induced disruption of cellular repolarization underlies electrocardiographic abnormalities that are diagnostic indicators of arrhythmia susceptibility.

Effects of cocaine on fixed-interval responding reinforced by the opportunity to run.

Rate-dependent drug effects have been observed for operant responding maintained by food, water, heat, light onset, electrical brain stimulation, shock-stimulus termination, and shock presentation. The present study sought to determine if the effects of cocaine on lever pressing maintained by the opportunity to run could also be described as rate dependent. Seven male Wistar rats were trained to respond on levers for the opportunity to run in a wheel. The schedule of reinforcement was fixed-interval 60 s, and the reinforcing consequence was the opportunity to run for 60 s. On this schedule, overall rates of responding were low, usually below six presses per minute, and pauses frequently exceeded the 60-s interval. Despite these differences, an overall scalloped pattern of lever pressing was evident for each rat. Doses of 1, 2, 4, 8, and 16 mg/kg cocaine were administered 10 min prior to a session. Only at the 16 mg/kg dose did the responding of the majority of rats change in a manner suggestive of a rate-dependent drug effect. Specifically, lower response rates at the beginning of the intervals increased and higher rates at the end of the intervals decreased, as indicated by the fact that slopes from the regression of drug rates on control rates decreased. These data provide tentative support for the generalization of rate-dependent effects to operant responding maintained by wheel running. Differences in the baseline performance maintained by wheel running compared to those for food and water point to the need for further experimentation before this effect can be firmly established.

A comparison of the effects of amphetamine, apomorphine and white noise on response switching in the rat

The effects of d-amphetamine, apomorphine and white noise on response switching in the rat were examined using a schedule of reinforcement which resulted in the subjects displaying a range of different probabilities of switching. The procedure was analogous to the use of a fixed interval schedule of reinforcement for examining the rate-dependent effects of drugs. d-Amphetamine (0.4-4.0 mg/kg) increased response switching in a manner dependent both upon the dose of drug and the baseline probability of switching. Apomorphine (0.01-0.3 mg/kg) increased switching in a manner which depended upon dose but which was independent of the baseline probability of switching. Neither drug increased response rate, although both drugs reduced response rate at the highest doses. In contrast, continuous white noise (85-105 dB) increased response rate without affecting switching. The results indicate that different activating stimuli may have qualitatively different effects on behaviour.

Effects of methsuximide on schedule-controlled responding in the pigeon

Acute and chronic effects of methsuximide (25, 50, 75, and 100 mg/kg), a succinimide with anticonvulsant properties, were examined in pigeons responding under a multiple fixed-ratio 50 fixed-interval 90-sec schedule of food delivery. The clearest acute effect of methsuximide was a substantial reduction in response rates under both components of the multiple schedule when the drug was administered at 100 mg/kg. Detailed analysis of the temporal distribution of responding under the fixed-interval failed to reveal rate-dependent drug effects. Tolerance appeared to develop to the effects of methsuximide when the drug was administered chronically.

Test of a model of antiarrhythmic drug action. Effects of quinidine and lidocaine on myocardial conduction.

The effects of quinidine and lidocaine on the maximum upstroke velocity (Vmax) of the ventricular myocardial action potential were compared with the effects predicted by a model over a wide range of driving rates, rhythm disturbances and holding potentials. These rate-, rhythm- and voltage-dependent effects were accurately predicted by the proposed model. The model was also able to predict several previously undocumented properties of the drugs: 1) If lidocaine decreases Vmax of a pulse train, the steady state is reached within a few action potentials. 2) The poststimulation recovery of Vmax in the presence of lidocaine or quinidine can occur in a multiexponential fashion, if the membrane potential is kept at the potential where both the fast (operating mainly at more negative membrane potentials) and the slow (operating at more positive potentials) recovery processes are operative. 3) Hyperpolarization markedly attenuates the rate-dependent drug effects. 4) Combinations of lidocaine and quinidine have a superadditive effect on the Vmax of early extrasystoles.

“Paradoxical” effects of psychomotor stimulant drugs in hyperactive children from the standpoint of behavioural pharmacology

Abstract Following a brief review of the effectiveness of psychomotor stimulant drugs in treating childhood hyperkinesis, the hypothesis of a “paradoxical” drug response is examined. The importance of three main factors, besides dosage, derived from consideration of studies in behavioural pharmacology. is assessed: rate-dependency; attention and stereotypy; and subjective and social effects. Rate-dependent effects of drugs in animals occur when the control rate of responding is related to the drug effect. A dose-dependent inverse relationship between control rate and drug effect is generally found within individual cases responding in operant situations, after treatment with stimulant drugs. This rate-dependency also occurs between organisms, such that those organisms with high control rates (for example, of operant behaviour, or spontaneous motor activity) show proportionately smaller increases in rate, or reductions in rate with amphetamine-like drugs. This article extends this form of analysis to hyperactive children in two populations receiving methylphenidate. In a third population, of normal adults and normal children, as well as hyperactive children, an inverse relationship between control rate of activity and the effects of 0.5 mg kg of d-amphetamine is found, accounting for about 38% of the variability. Therefore, the rate-reducing effects of psychomotor stimulants might arise in part as a consequence of the high rates of control responding in the hyperactive child, rather than from a unique “paradoxical” response. The effects of stimulants on “attention” in hyperactive children similarly might be predicted from studies on normal adults. Some evidence of an “over-focusing” of attention in hyperactive children treated with methylphenidate is presented in a test of cognitive flexibility. This lack of cognitive flexibility might be related to behavioural stereotypies found in animals and Man following stimulants. Finally, many of the subjective and social responses to stimulants in hyperactive children are not necessarily anomalous. The implications for clinical practice and for animal models of the hyperkinetic syndrome are discussed.

Mathematics relevant to rate-dependent drug effects

Abstract Changes in rates of responding after the administration of many drugs have been found to be related in a systematic manner to the rates of responding prior to drug administration. Recent controversy has centered on whether the drug effect should be measured in absolute or relative terms and on the limits imposed on the mathematical statement of the relationship by the maximum possible rate of responding, R m . The present paper suggests that both a redefinition of effect in absolute terms rather than in terms of change, and an incorporation of R m into the mathematical formulation, are unnecessary.

Rate dependent drug effects: Possible state dependency

Of 6 rats given extensive experience responding on a fixed-interval reinforcement schedule after injections of 1 mg/kg d-amphetamine, 3 showed significant rate dependent changes in responding when presession saline was substituted for d-amphetamine. Low rates were increased and high rates decreased by the change from d-amphetamine to saline, a result commonly found when d-amphetamine is first introduced. This indicates that rate dependent changes in fixed-interval responding may be state dependent phenomena.

Rate-dependent effects of drugs on the variable-interval behavior of rats.

Rate-dependent effects of drugs on the variable-interval behavior of rats.

Rate-dependent Effects of drugs. II. effects of some major tranquilizers on multiple fixed-ratio, fixed-interval schedule performance.

Rate-dependent Effects of drugs. II. effects of some major tranquilizers on multiple fixed-ratio, fixed-interval schedule performance.

ON THE FORM OF THE RELATION BETWEEN RESPONSE RATES IN A MULTIPLE SCHEDULE1

Three pigeons received training on multiple variable-interval schedules with brief alternating components, concurrently with a fixed-interval schedule of food reinforcement on a second key. Fixed-interval performance exhibited typical increases in rate within the interval, and was independent of multiple-schedule responding. Responding on the multiple-schedule key decreased as a function of proximity to reinforcement on the fixed-interval key. The overall relative rate of responding in one component of the multiple schedule roughly matched the overall relative rate of reinforcement. Within the fixed interval, response rate during one multiple-schedule component was a monotonic, negatively accelerated function of response rate during the other component. To a first approximation, the data were described by a power function, where the exponent depended on the relative rate of reinforcement obtained in the two components. The relative rate of responding in one component of the multiple schedule increased as a function of proximity to fixed-interval reinforcement, and often exceeded the overall obtained relative rate of reinforcement. The form of the function relating response rates is discussed in relation to findings on rate-dependent effects of drugs, chaining, and the relation between response rate and reinforcement rate in single-schedule conditions.

Drugs and punished responding. I. Rate-dependent effects under multiple schedules.

The effects of drugs were studied in pigeons whose responses were punished with electric shock during one component of a multiple fixed-interval 5-min fixed-interval 5-min schedule of food presentation. Most of the drugs analyzed for rate-dependent effects increased low rates of both punished and unpunished responding, while increasing higher rates less, or decreasing them; however, low rates of punished responding sometimes were increased more by pentobarbital, diazepam, and chlordiazepoxide than were matched rates of unpunished responding. In contrast, d-amphetamine and chlorpromazine usually increased low rates of unpunished responding more than matched rates of punished responding. These two drugs also decreased high rates of unpunished responding less than they decreased high rates of punished responding. Thus, the effects of drugs on punished responding depend on the control rate of punished responding; however, the rate-dependent effects of drugs on punished responding are not always the same as they are for unpunished responding.