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Some derivatives of the non-narcotic analgesic nefopam containing amidine and guanidine substituents have been prepared and their analgesic activity assessed by their ability to block the uptake of noradrenaline. The compounds have been shown to inhibit noradrenaline uptake but they also display possible α1 antagonist activity at higher concentration. An ester derivative was also active and was more selective as it did not exhibit α1 adrenoreceptor antagonism.

The a1- and a2-adrenoreceptor types are defined in terms of the selective binding of radioligands and their displacement from membrane preparations by a range of agonists and antagonists. Their distribution is described in brain, kidney, heart, and peripheral blood vessels. A group of centrally acting clonidine-like drugs are classified in terms of their selectivity for the a2-adrenoreceptors, and it is suggested that differences between a-adrenoreceptors in various tissues may allow development of highly selective a2-adrenoreceptor agonists for specific tissue sites such as the juxtaglomerular cells and brainstem nuclei controlling blood pressure. It is further suggested that the aim of treatment with such compounds is to modulate adrenergic activity while retaining normal homeostatic mechanisms. For this purpose, competitive agonists and antagonists are preferable to compounds which bind irreversibly to the receptor.

The muscarinic receptors in membranes prepared from guinea pig brain were studied using a radiolabeled antagonist, [3H]quinuclidinyl benzilate (QNB). The apparent dissociation constant of the QNB-receptor complex (Kd) was similar in all regions, but the concentration of receptors was highest in the striatum, cerebral cortex, and hippocampus and lowest in the cerebellum. Similar distributions have been reported for other species, although the concentration of receptors in guinea pig brain is higher than in other species. Acetylcholine inhibited QNB binding with a Hill coefficient of 0.4–0.6. The concentration of acetylcholine required to inhibit binding by 50% (I50) was lowest in the brain stem and more than 10 times higher in the hippocampus. Similar results have been reported for mouse brain. The activity of acetylcholinesterase was highest in the striatum, where the concentration of muscarinic receptors is highest, but did not vary greatly in other brain regions.

Phosphoinositide (PI) turnover, chronotropic and inotropic responses to α1-adrenoceptor activation, and α1-adrenoceptur density were studied in atria from rats with left ventricular myocardial infarction (LVMI) and noninfarcted rats. LVMI was produced after surgical ligation of the left coronary artery in 8-week-old Wistar rats. Rats were killed 4 weeks after this operation when rats with LVMI had developed significant hypertrophy of both ventricles and atria. Phenylephrine 0.1 4mUMto 1 mM. with propranolol 0.3 μM. produced a concentration-dependent increase in heart rate (HR) in right atria from noninfarcted rat hearts, and this response was significantly reduced in rats with LVMI. In electrically driven left atria, the concentration-dependent, phenylephrineinduced positive inotropic responses observed with propranolol added were also significantly impaired in rats with LVMI as compared with those of noninfarcted rats. In contrast, neither PI turnover in response to phenylephrine in the presence of propranolol nor α1-adrenoceptor density was reduced in rats with LVMI. These results suggest that the impaired α1-adrenoceptor–induced chronotropic and inotropic responses in atria from rats with LVMI are not due to downregulation of α1-adrenoceptors or to impaired activation of PI turnover after α1-adrenoceptor stimulation, but to impairment of one or more biochemical responses distal to PI hydrolysis or changes in coupling mechanisms other than hydrolysis of PIs.

The interaction between some examples of mononuclear and binuclear DNA-intercalating antitumor agents and alpha- and beta-adrenoceptors has been studied using radioligand-binding assays. Competition for 125I-BE 2254, [3H]rauwolscine, and (-)-[3H]dihydroalprenolol binding was used to assess affinity for alpha 1-, alpha 2-, and beta-adrenoceptor-binding sites, respectively. Two homologous series of alkyl-linked diacridines and diquinolines were found to interact poorly with beta-adrenoceptors, with only the largest members having appreciable affinity. By contrast, these compounds bind strongly and in a complex manner to alpha 1- and alpha 2-adrenoceptors. The affinity of diacridines for both alpha-adrenoceptor classes has a parabolic dependence on alkyl chain length with the hexyl and pentyl derivatives being the most potent at the alpha 1- (Ki = 11.5 +/- 2.3 nM) and alpha 2- (Ki = 143 +/- 26 nM) binding sites, respectively. The dependence of inhibition constants on linker chain length for the diquinolines is more complicated, with the ethyl- and heptyl-linked dimers having the greatest affinity for each alpha subclass. There is a nadir in affinity for the pentyl and butyl ligands and an increase in dissociation constant for octyl and longer homologues. Thus, the ethyl diquinoline has Ki values of 6.6 +/- 1.2 and 110 +/- 14 nM for the alpha 1- and alpha 2- adrenoceptors, respectively, and, correspondingly, the heptyl derivative has values of 39 +/- 4 and 51 +/- 1 nM. These findings are discussed with respect to a model of the alpha-adrenoceptor in which the radioligand-binding site is situated in a trench or cleft, surrounded by a flat surface bounded by walls. Daunomycin was found to have no affinity for adrenoceptors of any type and mitoxantrone similarly fails to interact with alpha 2- and beta-adrenoceptors, but binds to the alpha 1 subclass with an inhibition constant (Ki) of 3930 +/- 420 nM. Bisantrene also has no affinity for beta-adrenoceptors but binds to alpha 1- and alpha 2- adrenoceptors with Ki values of 145 +/- 24 and 2310 +/- 430 nM, respectively. Among the mononuclear acridine drugs studied, only nitracrine shows detectable interaction with beta-adrenoceptors (Ki = 760 +/- 50 nM). This compound, like bisantrene, has high affinity for the alpha 1-adrenoceptor (Ki = 131 +/- 17 nM) and moderate affinity for the alpha 2 subclass (Ki = 2180 +/- 500 nM).(ABSTRACT TRUNCATED AT 400 WORDS)

In a double-blind, placebo-controlled, parallel group study, 24 essential hypertensive subjects were randomised to receive either placebo or 2, 4, or 8 mg perindopril. Perindopril, its deesterified metabolite, perindoprilat, and perindoprilat glucuronide were separated with an ion-exchange resin and determined by a radioimmunoassay (RIA). Pharmacokinetic and pharmacodynamic parameters were estimated for 96 h after the first dose and after 4-week once-daily treatment. Perindopril peak levels were achieved in < 2 h after dosing with an elimination t1/2 of 1–2 h. Peak levels of perindoprilat were achieved more slowly, reaching a maximum level 5–8 h after dosing, and had an elimination t1/2 of 40 h. Levels of the perindopril glucuronide peaked ∼0.5 h later than perindopril, with an elimination t1/2 of ∼2 h. Perindopril, perindoprilat, and its glucuronide conjugate followed linear kinetics in the dose range of 2–8 mg, and there was no evidence of accumulation with chronic dosing. Perindopril 4 and 8 mg produced significant decreases in pre-dose blood pressure (BP) with chronic dosing, with maximal decreases occurring 5–7 h after dosing. Perindopril also produced a prolonged dose-dependent inhibition of plasma angiotensin-converting enzyme (ACE) activity that was maximum after 4 h and had not fully recovered by 48 h after a single dose.

The disposition of the potent antihypertensive drug clonidine has been poorly understood through the lack of a convenient and sensitive assay. A radioimmunoassay for clonidine has been developed and is capable of detecting as little as 10 pg of clonidine. 2,6-Dichlorophenyl-guanidine, a known metabolite of clonidine, did not cross-react with the antiserum whereas another metabolite, 4-hydroxyclonidine, was as potent as clonidine in displacing labeled clonidine from the antibody. However, a simple solvent extraction step before the radioimmunoassay selectively extracted clonidine from a mixture of clonidine and 4-hydroxyclonidine in alkaline plasma and this procedure permitted a specific assay for clonidine. The plasma levels of clonidine in rats after the administration of a hypotensive dose (100 microgram/kg i.v.) were determined by radioimmunoassay and these data indicated that the disposition of clonidine conforms to an open two-compartment, pharmacokinetic model. Clonidine rapidly accumulated in the brain as shown by the attainment of peak concentrations within 2 min of i.v. injection.

Seventeen patients who developed cardiogenic shock after acute myocardial infarction were treated with intravenously infused dopamine. In eight of these patients a stable blood pressure was attained, but oliguria or anuria persisted, and oral treatment with prazosin was instituted. Diuresis occurred in seven of these patients, but was followed by transient hypotension associated with a rapid rise in plasma prazosin levels in three. Four patients left hospital, and there were two long‐term survivors. Prazosin may be a useful adjunct to dopamine in the treatment of cardiogenic shock.

Summary The pharmacokinetics of clonidine and its relation to blood pressure response and side effects were studied after single oral doses of 75 µg, 150 µg and 250 µg in normotensive subjects. Following oral administration, the drug was absorbed rapidly after an initial lag time of 19–22 min and peak levels were reached between 2.4 and 2.9 h. Sampling over 48 h was necessary for accurate estimation of pharmacokinetic parameters. Post-peak plasma concentration declined in a monoexponential manner and the half-life of the elimination phase ranged from 9.0 to 15.1 h. Maximum plasma concentration (Cmax) and area under curve (AUC) increased proportionally with increasing doses. Clonidine produced significant reductions in the pulse rate and a dose dependent decrease in blood pressure. Clonidine (150 µg) also produced significant reductions in plasma catecholamine levels.

The antihypertensive effects of methyldopa and clonidine result from agonist activity at alpha adrenoceptor sites within the brain. Methyldopa is converted enzymatically to alpha-methylnoradrenaline in noradrenergic neurones in rat brain and replaces the natural transmitter, noradrenaline. Radioligand receptor studies show that alpha-methylnoradrenaline differs from noradrenaline in being much more selective (70 times) for the alpha2 subclass of adrenoceptors than noradrenaline and it is likely that the antihypertensive action of methyldopa results from selective activation of alpha2 adrenoceptors by alpha-methylnoradrenaline in the nucleus tractus solitarius and the anterior hypothalamus. Radioligand studies also show that clonidine is a selective alpha2 adrenoceptor agonist although it probably interacts with alpha] adrenoceptors at higher concentrations. With regard to a withdrawal syndrome after cessation of clonidine treatment, the cardiovascular and behavioural components can now be characterised in a rat model. The components include increases in basal blood pressure and heart rate, as well as increases in cardiovascular reactivity and also increases in rapid eye movement (REM) sleep, body shakes and tremor which is reminiscent of an opiate withdrawal syndrome. Increased central noradrenergic activity is involved in this syndrome and alpha1 and alpha2 adrenoceptors mediate opposing effects on the REM sleep rebound component.

The centrally acting α-adrenoceptor agonists clonidine, lofexidine, and S3341 vary in their selectivity for the α2-adrenoceptor subtype (S3341 > clonidine > lofexidine). The aim of this study was to examine the frequency and duration of rapid eye movement (REM) sleep episodes, and the cardiovascular changes associated with these episodes, during and after the continuous (10-day) infusion of saline (0.9 wt/vol), clonidine (10 μg/kg/h), lofexidine (10 μg/kg/h), or S3341 (50 or 100 μg/kg/h) in unrestrained, chronically instrumented rats. During a typical 24-h period, the saline infusion animals (n = 4) had 46 ± 3 REM sleep episodes of 2.4 ± 0.1 min duration. The majority of these episodes were associated with relatively minor phasic and tonic changes in arterial blood pressure and heart rate. During infusion, the three drugs markedly reduced the cardiovascular and behavioural signs of REM sleep. On cessation of the clonidine and lofexidine infusions, rebound increases (i.e., to values greater than controls) in the frequency and duration of REM sleep episodes were observed. These episodes were associated with markedly exaggerated fluctuations in arterial blood pressure and heart rate. The REM sleep rebound and associated cardiovascular responses were greater during the post-lofexidine infusion phase. A relatively minor rebound increase in total REM sleep time was observed after cessation of the 100-μg/kg/h, but not the 50-μg/kg/h, infusion rate of S3341. The cardiovascular responses during the REM sleep episodes were mildly exaggerated after cessation of the higher infusion rate of S3341. These results indicate that the magnitude of the REM sleep rebound (lofexidine > clonidine > S3341) may be inversely related to α2-adrenoceptor selectivity. As such, the α1-adrenoceptor activities of the clonidine and lofexidine infusions may underlie the subsequent rebound phenomenon.

We investigated the hypotensive effect and tissue distribution of prazosin (1 mg/kg i.v.) in conscious rats. This dose caused an immediate fall in blood pressure, associated with a nonsignificant increase in heart rate. Blood pressure then gradually rose to control over a 2-h period. Prazosin was measured in plasma and tissue samples by high-pressure liquid chromatography. Highest concentrations of drug were found in liver and kidney and the lowest in brain. The rate of decline of prazosin from plasma was similar to its decline from the tissues, indicating a rapid equilibrium. This rate of disappearance was also similar to the decay of the hypotensive response and suggests that prazosin in plasma may be in equilibrium with prazosin at its site of action.

The influence of cigarette smoking on plasma theophylline elimination half-life was examined in a group of elderly in-patients, to determine whether the modifying effect of cigarette smoking is also present in this age-group. The plasma theophylline half-life was determined in five non-smokers and seven smokers aged 66-88 years after administration of 320 mg of theophylline syrup. A wide range of plasma theophylline half-lives was observed in each group. However, there was a significant difference (P< 0.05) between the mean plasma theophylline half-life in smokers (5.41 h, SEM 0.69 h) and non-smokers (10.40 h, SEM 1.90 h). It was concluded that, although plasma theophylline half-life tends to be longer in this age-group, the modifying influence of cigarette smoking on plasma theophylline half-life can be demonstrated in the elderly, as well as in young adults.

Mitochondrial abnormalities are reported in four cases of phaeochromocytoma. These abnormalities include swelling and scant cristae, intramitochondrial dense bodies, septate-like junctions, intercristal fusion plus spheroidal bodies, and intramitochondrial rodlets. These structural mitochondrial changes are associated with reduction in activity of the mitochondrial enzymes, monoamine oxidase and succinic dehydrogenase.

We have measured the plasma concentrations of captopril and total disulfide conjugates of captopril after a 50 mg oral dose in 6 uraemic patients on maintenance dialysis and in 8 hypertensive subjects with normal renal function.

1. Studies with a sensitive radioenzymatic assay for plasma noradrenaline suggest there is a selective overactivity of the sympathetic nervous system in essential hypertension. 2. Serotonin turnover in the mesenteric vessels is approximately twice that of noradrenaline and it is suggested that serotonin may interact with noradrenaline to maintain vascular resistance. 3. Methodology which allows the study of local sympathetic turnover in nuclei of the central nervous system and in peripheral blood vessels is described. This approach has been used to study non-innervated sympathetic turnover observed in phaeochromocytoma.

1. The activities of the enzymes involved in catecholamine synthesis, tyrosine hydroxylase, aromatic amino acid decarboxylase and dopamine ß-hydroxylase, were markedly enhanced in homogenates of phaeochromocytoma compared with human adrenal medulla homogenates measured under optimum substrate concentrations. 2. It was demonstrated in fresh tumour slices that the rate of formation of dopamine (3,4-dihydroxyphenethylamine) from tyrosine was much slower than the rate of formation of dopamine from dopa (3,4-dihydroxyphenylalanine) (suggesting that tyrosine hydroxylase was the rate-limiting enzyme in noradrenaline synthesis in phaeochromocytoma) and that the tyrosine hydroxylation step was still susceptible to end-product inhibition by catecholamines. 3. It is suggested that catecholamine overproduction in phaeochromocytoma is due to the increased activities of catecholamine synthetic enzymes rather than to an insensitivity of tyrosine hydroxylase to end-product inhibition. 4. The activities of the enzymes involved in catecholamine catabolism, monoamine oxidase and perhaps catecholamine O-methyltransferase, were reduced in phaeochromocytoma. This finding provides a biochemical basis for the previous published observations of nonexocytotic release of catecholamines in these tumours. Thus excess amounts of newly synthesized noradrenaline which cannot be stored in the filled catecholamine storage vesicles may not be degraded owing to the reduced monoamine oxidase activity and could diffuse from the phaeochromocytoma into the circulation.

1. The question of cellular localizations of monoamine oxidase and catechol‐o‐methyl transferase, enzymes involved in the metabolism of noradrenaline, has been investigated by following the changes in the enzyme activity of the smooth muscle of the cat nictitating membrane after sympathetic denervation. Any falls in enzyme activity coinciding with the time course for degeneration of the sympathetic nerve endings (2–3 days) can reasonably be ascribed to a former localization of the enzyme within the sympathetic nerve.

The concentrations of glucose were elevated whereas those of insulin were decreased in the sera of normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive (SH) rats on day 10 of a continuous subcutaneous infusion of Clonidine (10 μg kg−1h−1for 10 days). Fifteen to 18 h after cessation of infusion, the glucose levels of both strains had fallen to below those of the respective controls whereas the insulin levels remained suppressed. As such the hypoglycaemia may be related to the general increase in metabolic requirements associated with the post-infusion ‘withdrawal syndrome’.

The effects of chronic treatment with two benzodiazepines were studied on kindled amygdaloid seizures in rats. Clobazam (4 mg kg−1), clonazepam (0.3 mg kg−1) or vehicle (1 mL kg−1) was administered, by intraperitoneal injection, to fully kindled rats twice daily for nineteen days. Each rat was electrically stimulated 30 min after the morning dose on alternate days of treatment. Tolerance developed rapidly to the anticonvulsant effects of clobazam after only three days of treatment, following which only a small residual protection was maintained. Tolerance to clonazepam developed gradually over the course of the experiment, although this effect was relatively minor.

In rats the continuous infusion of clonidine (10 μg kg−1h−1) produces a relative hyperthermia of approximately 0·6 °C during the light phases but not the dark phases of a 10 day infusion period. It appears that the relative hyperthermia results from a clonidine infusion-induced attenuation of the fall in body temperature which normally occurs at the onset of the light phases.

A sensitive assay is described for quantitating plasma captopril levels. Captopril is an orally active inhibitor of angiotensin-con-verting enzyme. Blood from patients taking this drug was collected into tubes containing edetate and ascorbic acid, and the plasma was separated by centrifugation. After addition of an internal standard, the plasma was deproteinized and the supernate was adjusted to pH 6.5. N-(1-Pyrene)-maleimide was added to derivatize captopril and an internal standard to fluorescent adducts. These derivatives then were extracted into ethyl acetate-benzene (1:1) and separated from other derivatized thiols by high-performance liquid chromatography. The sensitivity of the assay was 150 pmoles/ml. Preliminary pharmacokinetics were obtained in patients taking captopril chronically for essential hypertension. After administration of 100mg of captopril to patients who had fasted overnight, the plasma levels rose rapidly; peak levels were obtained at ˜37 min. Thereafter, the plasma levels declined rapidly, and the terminal half-life was ˜40 min. In these patients, the time course of the plasma levels did not reflect changes in blood pressure, so there appeared to be no direct relationship between plasma levels of the parent compound and blood pressure changes.