Site selectivity of competitive antagonists for the mouse adult muscle nicotinic acetylcholine receptor.

The muscle-type nicotinic acetylcholine receptor has two nonidentical binding sites for ligands. The selectivity of acetylcholine and the competitive antagonists (+)-tubocurarine and metocurine for adult mouse receptors is known. Here, we examine the site selectivity for four other competitive antagonists: cisatracurium, pancuronium, vecuronium, and rocuronium. We rapidly applied acetylcholine to outside-out patches from transfected BOSC23 cells and measured macroscopic currents. We have reported the IC(50) of the antagonists individually in prior publications. Here, we determined inhibition by pairs of competitive antagonists. At least one antagonist was present at a concentration producing > or =67% receptor inhibition. Metocurine shifted the apparent IC(50) of (+)-tubocurarine in quantitative agreement with complete competitive antagonism. The same was observed for pancuronium competing with vecuronium. However, pancuronium and vecuronium each shifted the apparent IC(50) of (+)-tubocurarine less than expected for complete competition but more than expected for independent binding. The situation was similar for cisatracurium and (+)-tubocurarine or metocurine. Cisatracurium did not shift the apparent IC(50) of pancuronium or vecuronium, indicating independent binding of these two pairs. The data were fit to a two-site, two-antagonist model to determine the antagonist binding constants for each site, L(alphaepsilon) and L(alphadelta). We found L(alphaepsilon)/L(alphadelta) = 0.22 (range, 0.14-0.34), 20 (9-29), 21 (4-36), and 1.5 (0.3-2.9) for cisatracurium, pancuronium, vecuronium, and rocuronium, respectively. The wide range of L(alphaepsilon)/L(alphadelta) for some antagonists may reflect experimental uncertainties in the low affinity site, relatively poor selectivity (rocuronium), or possibly that the binding of an antagonist at one site affects the affinity of the second site.

Anesthesia blocks nonshivering thermogenesis in the neonatal rabbit.

Nonshivering thermogenesis (NST) is a normal physiological response of the neonate to cold exposure, characterized by increased blood flow to metabolically active brown fat stores. It is standard practice during neonatal surgery to warm the ambient environment in order to avoid consuming vital energy stores. While NST has been well-studied in the neonate, the response during anesthesia and paralysis has not been fully characterized. Rabbit pups (aged 1 to 7 days) were randomized into several groups. The experimental groups consisted of animals mechanically ventilated and administered either metocurine, pancuronium, curare, fentanyl, nitrous oxide (N2O), or halothane. The controls were spontaneously breathing animals. Oxygen consumption (VO2), an index of metabolic activity, was measured at thermoneutrality (39 degrees C) and after cold exposure (25 degrees C). Control and metocurine animals had a significant increase in VO2 in response to cold exposure. The increase in VO2 was not noted in animals that received curare, pancuronium, fentanyl, N2O, or halothane. To test the effect of anesthetic withdrawal during cold exposure on VO2, additional series of animals were studied. One group received continuous halothane throughout the period of cold exposure; the other had cessation of the halothane during cold exposure. Both groups were rewarmed subsequently. The animals that had withdrawal of halothane during cold exposure had a marked and significant increase in VO2 compared with the control group (continuous halothane). VO2 returned to near-baseline levels upon rewarming. The authors conclude that many commonly used anesthetic and paralyzing agents inhibit the thermogenic response to cold exposure. However, cessation of anesthesia (halothane) in a cold environment results in a marked increase in metabolic activity.

Molecular dissection of subunit interfaces in the acetylcholine receptor: identification of residues that determine curare selectivity.

The acetylcholine receptor from vertebrate skeletal muscle is a transmembrane channel that binds nerve-released acetylcholine to elicit rapid transport of small cations. Composed of two alpha subunits and one beta, one gamma, and one delta subunit, the receptor is a cooperative protein containing two sites that bind agonists, curariform antagonists, and snake alpha-toxins. Until recently the two binding sites were thought to reside entirely within each of the two alpha subunits, but affinity labeling and expression studies have demonstrated contributions by the gamma and delta subunits. Affinity labeling and mutagenesis studies have identified residues of the alpha subunit that contribute to the binding site, but the corresponding gamma- and delta-subunit residues remain unknown. By making gamma-delta chimeras and following the nearly 100-fold difference in curare affinity for the two binding sites, the present work identified residues of the gamma and delta subunits likely to be near the binding site. Two sets of binding determinants were identified in homologous positions of the gamma and delta subunits. The determinants lie on either side of a disulfide loop found within the major extracellular domain of the subunits. This loop is common to all acetylcholine, gamma-aminobutyrate, and glycine receptor subunits.

Comparison of lowest energy conformations of dimethylcurine and methoxyverapamil: evidence of ternary association of calcium channel, Ca2+, and calcium entry blockers.

Verapamil and dimethylcurine are Ca2+ entry blockers of essentially different chemical structures which presumably bind to the same arylalkylamine receptor of the L-type Ca channel. A systematic conformational analysis of methoxyverapamil (D-600) and dimethylcurine has been carried out using a molecular mechanics method. The lowest minimum-energy conformations of D-600 are predisposed to chelate Ca2+ by four oxygen atoms of the stacked methoxyphenyl moieties. Comparison of the lowest energy conformations of D-600-Ca2+ and dimethylcurine revealed a similar spatial disposition of cationic groups and methoxyphenyl moieties in the two compounds. A three-dimensional model of arylalkylamine receptor was suggested which incorporates two nucleophilic areas of the Ca channel. Dimethylcurine binds to these areas by its quaternary amine functions, whereas D-600 does so by amine function and via coordinated Ca2+. The results support the hypotheses on ternary complex formation between the ligands of Ca channel, their receptors, and Ca2+.

The influence of respiratory-induced acid-base changes on the action of non-depolarizing muscle relaxants in rats.

The influence of respiratory-induced acid-base changes on the action of non-depolarizing muscle relaxants was investigated using the rat phrenic nerve-hemidiaphragm preparation. Changes in pH were induced by changes in the CO2 concentration aerating Krebs' solution. In the absence of muscle relaxants, an increase in CO2 from 5% to 7.5% decreased (P less than 0.01) indirectly elicited twitch tension by 5.4 +/- 0.7% (mean +/- SEM), while a decrease in CO2 from 5% to 2.5% increased (P less than 0.01) twitch tension by 2.3 +/- 0.7%. With a change in CO2 from 2.5% to 7.5%, partial neuromuscular blockade produced by d-Tc or vecuronium was augmented (P less than 0.01), while that produced by metocurine, pancuronium, or alcuronium was reduced (P less than 0.01). With the change in CO2 from 7.5% to 2.5%, the neuromuscular blockade produced by d-Tc or vecuronium was reduced (P less than 0.01), while that produced by metocurine, pancuronium, or alcuronium was augmented (P less than 0.01). Dose-response study showed that 2.5% CO2 shifted the dose-response curves for d-Tc and vecuronium to the right (P less than 0.01) from those with 5% CO2, whereas 7.5% CO2 shifted them to the left (P less than 0.05). In contrast, neither 2.5% CO2 or 7.5% CO2 significantly shifted the dose-response curves for metocurine or pancuronium from those with 5% CO2. Their dose-response curves with 2.5% CO2 were to the left, instead of to the right, of those with 7.5% CO2 (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

The neuromuscular blocking effect of trimetaphan alone and in combination with different non-depolarizing muscle relaxants in the rat.

The effect of trimetaphan alone and in combination with pancuronium, tubocurarine or metocurine (dimethyl tubocurarine) has been examined on the rat phrenic nerve diaphragm preparation. Trimetaphan alone produced neuromuscular blockade in an all-or-none fashion once a concentration of between 2.39 X 10(-4) and 3.00 X 10(-4) M had been exceeded. Concentrations of trimetaphan below this threshold produced a dose-dependent potentiation of all three non-depolarizing relaxants studied. This potentiation was equal for tubocurarine and metocurine, but less for pancuronium.

Suppression of shivering decreases oxygen consumption and improves hemodynamic stability during postoperative rewarming.

Thirty-three patients undergoing elective myocardial revascularization were prospectively randomized into two study groups (Group S and Group P) to permit evaluation of the effects of shivering on oxygen consumption per minute (VO2), carbon dioxide production per minute (VCO2), and hemodynamic performance. Group S was allowed to shiver during the postoperative rewarming period, and Group P received hourly injections of pancuronium bromide and Metubine (metocurine) sulfate with sedation to block the shivering response. Group S demonstrated significantly higher increases in VO2 and VCO2, lower systolic blood pressure and mixed venous oxygen saturation, and a greater use of inotropic support than the patients in Group P. Suppression of the shivering response minimized increases in VO2 and VCO2, improved hemodynamic stability, and resulted in a decreased need for inotropic support.

Cardiovascular and neuromuscular responses to high-dose pancuronium-metocurine in pediatric burned and reconstructive patients.

The efficacy of the combined use of pancuronium and metocurine (Pm-MTC) in high doses to produce rapid-onset muscle paralysis was evaluated in 15 patients with acute burns and 18 recovered burned patients scheduled for reconstructive surgery. Two and three times the previously determined ED95 of the combination for each group was used. (ED95 for Pm-MTC combination is 0.032/0.129 mg/kg for acute burns and 0.013/0.051 mg/kg for reconstructive patients.) Doubling ED95 produced 95% paralysis in 3.1 +/- 0.9 min in acutely burned children and in 4.3 +/- 0.7 min in reconstructive children (mean +/- SEM). These onset times were not significantly different from each other. Tripling the ED95 of the combination in burned children reduced the onset time to 1.3 +/- 0.14 min, but this was not significantly different from 2 X ED95 onset time in burned patients. The administration of 3 X ED95 to the reconstructive group, however, resulted in a significantly more rapid onset time of 1.8 +/- 0.4 min compared with 2 X ED95 in the same population. With 3 X ED95 the onset times between burn and reconstructive patients were not significantly different. Time for recovery of twitch to 25% of control twitch height (75% twitch depression) was significantly prolonged in burned patients compared with reconstructive patients for equipotent doses administered. Although the occasional patient showed prominent changes in heart rate and blood pressure, overall cardiovascular stability was impressive.(ABSTRACT TRUNCATED AT 250 WORDS)

Cardiovascular effects of non-depolarizing neuromuscular blockers in patients with coronary artery disease.

To compare haemodynamic responses associated with equipotent doses of muscle relaxants and high dose fentanyl (50 micrograms X kg-1), 40 non-hypertensive patients who were receiving beta adrenergic and calcium channel blocker therapy and undergoing coronary bypass surgery were randomized to four study groups receiving the following: (1) atracurium: 0.4 mg X kg-1, (2) pancuronium: 0.12 mg X kg-1, (3) vecuronium: 0.12 mg X kg-1, or (4) pancuronium-metocurine mixture: (0.4 mg + 1.6 mg X ml-1):1 ml/10 kg. Neuromuscular blockers were injected with fetanyl at induction. Haemodynamics were recorded with the patients awake (baseline), at two minutes post-induction, and at two and five minutes after intubation. Pancuronium was the only drug associated with significant increases in HR; no other significant changes occurred within each group when compared to their respective baseline haemodynamics. HR increased more after induction with pancuronium when compared to atracurium (23 vs. 4 per cent, p less than 0.05) and to vecuronium (23 vs. 2 per cent, p less than 0.05), and when compared to vecuronium after intubation (29 vs. 7 per cent, p less than 0.05). The pancuronium-metocurine mixture caused tachycardia which was less than, though not significantly different than with pancuronium; however, HR returned to baseline by five minutes with the mixture, but remained elevated with pancuronium (3 vs. 18 per cent, p less than 0.05). SVR fell more on induction with atracurium when compared to vecuronium (-18 vs. 1 per cent, p less than 0.05). These changes in HR or SVR were not accompanied by ECG signs of ischaemia.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of muscle movement on the electroencephalogram during anesthesia with alfentanil.

Using aperiodic analysis, we examined the impact on the electroencephalogram (EEG) of muscle activity from opiate-induced rigidity with alfentanil. We compared two groups of patients, one receiving alfentanil with neuromuscular blocking agents and the other group receiving no relaxants. The alfentanil-induced muscle rigidity exerted a noticeable effect on the EEG, with a moderate effect on total power at 1 Hz; a marked effect on the total number of waves, cumulative percent power at 3 Hz, and average power at 17 to 19 Hz; and a striking effect on F90, the frequency below which 90% of the power resides. The presence of electromyographic (EMG) noise in the EEG consistently altered the variables derived from the EEG, so that anesthetic depth appeared less than it actually was. This was true in spite of the fact that we gave slightly more alfentanil in the group not receiving a relaxant. Although the observed muscle activity was greater than that usually seen clinically, and may have differed qualitatively, the results do serve as a warning that muscle noise can interfere with the EEG. Currently, there is no computerized technique that will reject or account for this noise, and we must depend on observation to recognize the EMG patterns within the EEG, either with the raw recording or with a detailed analysis (such as aperiodic analysis), and to compensate for this noise if possible. Techniques that average the EEG or that present a single number have difficulty providing this information. These results do not detract from the usefulness of the EMG contained in EEG recordings as a supplementary or complementary indicator of anesthetic lightness.

Interaction among agents that block end-plate depolarization competitively.

The influence of gallamine, metocurine, pancuronium, and tubocurarine on depolarization of a mammalian muscle end-plate region was examined to determine whether the antagonists given in combination exerted a degree of block consistent with the simple classical competitive model. Depolarization was produced by carbachol in isolated guinea pig lumbrical muscles and recorded by the moving fluid electrode technique. The dose-response values obtained were fitted to a regression embedded in a split plot factorial experimental design such as both to control and to measure effects of variation among preparations, order of administration, time, and level of block. Of the six possible pairings of the four drugs, four showed the simple additivity expected from a competitive mechanism, while two (pancuronium plus metocurine and metocurine plus gallamine) showed potentiation beyond additivity. In these latter two pairs the combination shifted the carbachol dose-response curve, respectively, 41 and 21% further than predicted from the classical model. The significance of this deviation in the light of alternative receptor models is discussed, and a model consistent with the observed results is outlined.

Adverse effects of pancuronium during high-dose fentanyl anesthesia for coronary artery bypass grafting.

Using a randomized double-blind protocol, the authors prospectively compared three nondepolarizing muscle relaxants with respect to their influence on hemodynamics and on the electrocardiogram. Thirty-three patients undergoing elective coronary artery bypass grafting (CABG) with high-dose (100 micrograms/kg) fentanyl anesthesia were studied. Patients received 1.5 X ED95 of either pancuronium (n = 12), metocurine (n = 9), or a metocurine-pancuronium combination (4:1 ratio by weight) (n = 12) for muscle relaxation. Heart rate and rate pressure product (RPP) were significantly higher postinduction in the pancuronium group. Myocardial ischemia, indicated by new ECG ST-segment depression occurred significantly more frequently, and exclusively, in the pancuronium group. The authors' data suggest that since pancuronium is associated with tachycardia and an increased incidence of myocardial ischemia, it is best avoided in patients with severe coronary artery disease undergoing CABG with high-dose fentanyl. Either metocurine or the metocurine-pancuronium combination provides greater hemodynamic stability, without precipitating myocardial ischemia, and can be safely and effectively substituted for pancuronium.

Potentiation of the combination of pancuronium and metocurine by halothane and isoflurane in humans with and without renal failure.

Dose-response relationships for a 1:4 weight ratio-mixture of pancuronium and metocurine were studied during inhalational anesthesia with halothane and isoflurane in patients with and without renal failure. The time for recovery from 10 to 20% of control thumb twitch tension also was determined. In subjects with normal renal function, relaxant doses required for 95% twitch height suppression (ED95) were 50% of those predicted by simple addition of effects when used with a balanced anesthetic technique, 37% of predicted when used with 1.3 MAC halothane, and 25% of predicted when used with 1.3 MAC isoflurane (P less than 0.05). In subjects with renal failure, ED95 values for the combination were 40% of predicted when used with 1.2 MAC halothane and 45% of predicted when used with 1.2 MAC isoflurane (NS). For relaxants used singly in renal failure, pancuronium alone was slightly enhanced by 1.2 MAC halothane (85% of predicted), while 1.1 MAC isoflurane reduced the ED95 to 57% of predicted (P less than 0.05). Similar results were obtained for metocurine alone when used in renal failure (77 and 58% of predicted when used with halothane and isoflurane, respectively) (NS). Predicted values are published results for balanced anesthesia in normals. Recovery times were prolonged twofold in renal failure (P less than 0.05). Thus, the combination of pancuronium and metocurine is synergistic to the same degree in normals and in renal failure patients, but the total blockade produced by the combination is enhanced by halothane and isoflurane only in normals.

Facilitation of rapid endotracheal intubations with divided doses of nondepolarizing neuromuscular blocking drugs.

The authors sought to determine whether prior administration of a small, subparalyzing dose of nondepolarizing muscle relaxant would shorten the onset time of an intubating dose of muscle relaxant. Initially, in 60 anesthetized patients, twitch response of adductor pollicis to ulnar nerve stimulation was studied after a small dose of pancuronium 0.015 mg . kg-1, metocurine 0.03 mg . kg-1, or d-tubocurarine 0.04 mg . kg-1, followed 3 min later by pancuronium 0.08 mg . kg-1 or atracurium 0.4 mg . kg-1 administered iv. After 60 s, the minimum neuromuscular block, in all patients was 79.0 +/- 5.0%. A 95% depression or twitch tension occurred between 59.1 +/- 5.3 and 86.1 +/- 5.9 s. In another 60 patients, intubating conditions under similar regimen were studied, except the small dose of muscle relaxant was given immediately prior to induction of anesthesia. At the end of 60 s, good to excellent intubating conditions were present in 100% of the patients following the second dose of pancuronium and in 83% of the patients following atracurium. In 17% of the patients, after atracurium intubating conditions were fair. When nondepolarizing neuromuscular blocking drugs are administered in divided doses, neuromuscular blockade adequate for endotracheal intubation is achieved in less than 90 s. This facilitates rapid endotracheal intubation in a time comparable to using succinylcholine, without undesirable effects of the depolarizing neuromuscular blocking drugs.

Resistance to competitive neuromuscular blocking agents in burn patients: a review.

The quantitative and qualitative aspects of the phenomenon of resistance to competitive (non-depolarizing) neuromuscular blocking agents in burn patients are described. The correlates and temporal features of this resistance are discussed, in addition to therapeutic approaches and the possible mechanisms underlying the resistance.

The dose response effect of long-acting nondepolarizing neuromuscular blocking agents in children.

Cumulative dose-response curves were constructed for pancuronium, metocurine, d-tubocurarine and gallamine in 56 children anaesthetized with thiopentone, N2O/O2 and narcotic. The dose response curves of the four relaxants did not deviate significantly from parallelism. The effective dose causing 95 per cent depression of the twitch at 0.1 Hz was: pancuronium 0.08 mg X kg-1, metocurine 0.34 mg X kg-1, d-tubocurarine 0.6 mg X kg-1, and gallamine 3.4 mg X kg-1. Thus, pancuronium is 40 times more potent than gallamine, while metocurine and d-tubocurarine are seven and four times more potent than gallamine. The recovery of twitch height from 5-25 per cent of control for pancuronium (15.6 +/- 1.7 min) was significantly faster (p less than 0.01) than metocurine (27.3 +/- 1.9 min), d-tubocurarine (32.2 +/- 4.8 min), or gallamine (30 +/- 3.3 min). Compared to studies in adults, the present data indicate that children have a tendency (statistically not significant) to require more relaxant and recover more quickly than adults.

Potentiation of neuromuscular blockade using a combination of pancuronium and dimethyltubocurarine. Studies in children following acute burn injury or during reconstructive surgery.

The neuromuscular effects of the combined administration of pancuronium and dimethyltubocurarine were evaluated in children undergoing reconstructive surgery (n = 6), or the skin grafting of acute burn wounds: body surface area (BSA) burns less than 40% (n = 5); and BSA burns greater than 40% (n = 6). A dose of pancuronium 0.005 mg kg-1 was considered to be equipotent with dimethyltubocurarine 0.02 mg kg-1, and each dose was defined as being equal to 1 relaxant equivalent (RE). Incremental doses of the combination of pancuronium and dimethyltubocurarine were administered until a 95% depression of twitch height was achieved (ED95). These results were compared with previously published data for pancuronium and dimethyltubocurarine alone. The mean ED95 of the combination in the control population (reconstructive surgery) was 5.1 RE, compared with 10.8 RE and 10.0 RE for pancuronium and dimethyltubocurarine , respectively, administered alone. In the acutely burned population, the mean ED95 for the combination was 9.9 and 15.9 RE, respectively, compared with 26 RE for pancuronium alone in burned patients. In all groups of patients, significantly less total drug was required when the combined therapy was used.

Cardiovascular effects of nondepolarizing relaxants employed for pretreatment prior to succinylcholine.

A pregnant woman with severe pre-eclampsia experienced a hypertensive crisis following a pretreatment dose (20 mg) of gallamine. That episode initiated a study to determine the cardiovascular effects of non-depolarizing muscle relaxants in 58 nonobese, ASA physical status I and II adults. Subjects were assigned randomly to one of five treatment groups as follows: gallamine (0.29 mg X kg-1), d-tubocurarine (0.04 mg X kg-1), metocurine (0.014 mg X kg-1), pancuronium (0.007 mg X kg-1), or normal saline (control). Baseline measurements of systolic, diastolic, mean arterial pressure, heart rate (HR) and rate pressure product (calculated RPP) were recorded at one-minute intervals while electrocardiogram, lead II, was recorded continuously. Statistically significant increases occurred in HR at minutes 2, 3 and 4; RPP at minutes 3 and 4; and per cent change in HR at minutes 2, 3 and 4 following gallamine pretreatment. The rise in RPP was predominantly due to the elevation in HR. These results suggest that even modest doses of gallamine should be avoided in clinical situations where lability of cardiovascular dynamics can be anticipated.

Pancuronium, unlike other nondepolarizing relaxants, retains potency at hypothermia.

The authors studied the pharmacodynamics of four nondepolarising relaxants, d-tubocurarine (dTC), pancuronium, metocurine, and gallamine, at 25 degrees C, 31 degrees C, and 37 degrees C. The rat phrenic nerve-hemidiaphragm preparation with vascular perfusion was used for these investigations. For each drug at each temperature, a dose-response curve for twitch depression was constructed. ED50 values were calculated using probit-log dose regression. dTC, metocurine, and gallamine each demonstrated a near twofold increase in ED50 at 25 degrees C compared with 37 degrees C. No such relationship was apparent with pancuronium. In addition, the slopes of the dose-response curves were analyzed for effects due to temperature or drug. Slopes were not influenced by temperature; however, the slopes for metocurine and dTC were lower than those for pancuronium and gallamine. The authors conclude that in the rat, pancuronium retains potency at hypothermia, whereas the other relaxants decrease potency. In addition, metocurine and dTC exhibit less steep dose-response curves under these experimental conditions.