Pharmacological characterization of SR 47436, a new nonpeptide AT1 subtype angiotensin II receptor antagonist.

SR 47436, 2-n-butyl-3-[(2'-(1H-tetrazol-5-yl)-biphenyl-4-yl) methyl]-1,3-diaza-spiro[4,4]non-1-en-4-one, is a new potent and selective AT1 angiotensin II (AII) receptor antagonist. It competitively inhibited [125I]AII binding to AT1 subtype receptors in rat liver membranes (IC50 = 1.7 nM) and did not interact with AT2 subtypes in rat adrenal cortical membranes. In rabbit aorta, SR 47436 inhibited contractions induced by 10 nM AII (IC50 = 4.0 nM) and shifted AII contractile response curves to the right in a parallel fashion, without total recovery of the maximal response. The potency of SR 47436 was higher than that of the lead compound, 2-n-butyl-4-chloro-5-hydroxymethyl-1-[(2'-(1H-tetrazol-5- yl)biphenyl-4-yl)methyl]imidazole (DuP 753) (rat liver binding: IC50 = 16 nM; rabbit aorta: IC50 = 26 nM), and equivalent to saralasin (IC50 = 1.8 and 2.7 nM, respectively). The high specificity of SR 47436 was demonstrated by its lack of activity (IC50 > 10 microM) on various other receptors, ionic channels and antiports and rabbit aorta contracted by norepinephrine and KCl, and its lack of inhibition of renin and converting enzyme. In conscious rats, SR 47436 as well as DuP 753 (0.1 to 3 mg/kg, i.v., and 0.3 to 30 mg/kg, p.o.) antagonized the AII-pressor response in a dose-related manner. In conscious dogs, SR 47436 (1-10 mg/kg, p.o.) was a more potent antagonist of the AII pressor response than DuP 753. In conscious chronically implanted cynomolgus monkeys, SR 47436 antagonized the AII-pressor response at 1 mg/kg (89% i.v. and 66% p.o.) much more strongly than DuP 753 at 10 mg/kg (83% i.v. and 20% p.o.).(ABSTRACT TRUNCATED AT 250 WORDS)

Biochemical and pharmacological properties of SR 46349B, a new potent and selective 5-hydroxytryptamine2 receptor antagonist.

A new potent, selective and p.o. active serotonergic [5-hydroxytryptamine (5-HT2)] receptor antagonist, SR 46349B [trans, 4-([3Z)3-(2-dimethylaminoethyl)oxyimino-3(2-flurophenyl++ +)propen-1-yl]phenol hemifumarate) has been characterized by a series of "in vitro" and "in vivo" methods. Based upon binding studies with 5-HT2 receptors in rat brain cortical membranes and blockade of 5-HT-induced contractions in isolated tissues (rabbit thoracic aorta, rat jugular vein, rat caudal artery, rat uterus and guinea pig trachea), SR 46349B showed high affinity for 5-HT2 receptors. Furthermore, SR 46349B displayed moderate affinity for the 5-HT1C receptor and had no affinity for the other 5-HT1 subclass (5-HT1A, 5-HT1B or 5-HT1D), dopamine (D1 or D2), "alpha" adrenergic (alpha-1 or alpha-2), sodium and calcium channel and histamine (H1) receptors. It did not interact with histamine (H1), alpha-1 adrenergic and 5-HT3 receptors in smooth muscle preparations. No inhibition of the uptake of norepinephrine, dopamine or 5-HT was seen. Based upon blockade of pressor responses to 5-HT in pithed rats and in vivo binding studies in mice, SR 46349B was found to be a potent and p.o. active 5-HT2 receptor antagonist with a relatively long duration of action. Behavioral experiments, including mescaline- and 5-hydroxytryptophan-induced head twitches and learned helplessness, as well as sleep-waking cycle and EEG spectral parameter studies, indicated that SR 46349B has a classical 5-HT2 psychopharmacological antagonist profile.

Central alpha 1-adrenergic and opiate systems in the control of the vagal tone in normotensive and spontaneously hypertensive rats.

In anesthetized, normotensive beta-blocked rats, the alpha 1-adrenoceptor blocking drug, AR-C 239 (300 micrograms/kg, i.v.) induced a bradycardic effect related to a central increase in the vagal tone. This bradycardia was inhibited by previous administration of naloxone, intravenously (1 mg/kg) or centrally (100 micrograms/kg, i.c.) injected. Naloxone, by itself did not change the heart rate. In brainstem membranes from normotensive rats, AR-C 239 did not influence the stereoselective binding of [3H]naloxone. In spontaneously hypertensive (SH) rats, naloxone peripherally or centrally administered did not influence the activation of the vagal tone induced by AR-C 239, in beta-blocked animals. These results suggest the possible involvement of opiate release in the AR-C 239-induced vagal bradycardia, in normotensive rats. They also afford new arguments for the existence of close interactions between central alpha-adrenergic and opiate systems in the cardiovascular regulation. The possible participation of kappa-receptors in this effect is discussed. In addition, such an opiate mechanism triggered by central alpha 1-adrenoceptor blockade seems to be either absent or inactive in SH rats.

Further investigations on the alpha 1-adrenoceptor blocking properties of AR-C 239 in rats.

AR-C 239, a new alpha-adrenoceptor blocking drug, appears to act selectively on alpha 1 sites in rats. At peripheral sites, this drug did not change the tachycardia induced by spinal sympathetic outflow stimulation in pithed rats, and did not antagonize the inhibitory effects of clonidine on this preparation. In addition, AR-C 239 showed predominant alpha 1-adrenoceptor blocking properties in the bisected rat vas deferens preparation. AR-C 239 did not prevent or reverse the centrally mediated hypotensive and bradycardic actions induced by clonidine, in intact animals. In conclusion, AR-C 239 seems to be a very useful tool for the characterization of peripheral and central alpha 1-adrenoceptors, in this animal species.

Multiple sites for the cardiovascular actions of fentanyl in rats.

We studied the cardiovascular effects of intravenously administered fentanyl in normotensive rats anesthetized with pentobarbital and artificially ventilated. Fentanyl induced an immediate and short-lasting fall in blood pressure and heart rate by an action on opiate receptors localized at vagal nerve endings. Bilateral vagotomy suppressed these effects. The bradycardia, suppressed by bilateral vagotomy and reduced by previous administration of atropine, seemed to be due to a vagovagal reflex. Inhibition of the sympathetic outflow may also occur, because in pithed rats fentanyl failed to lower blood pressure. This masks a direct central stimulation of sympathetic outflow, because in bilaterally vagotomized rats fentanyl induced an alpha-adrenoceptor-blocking drug-sensitive hypertension which was insensitive to adrenalectomy. In addition, stimulation of cardiac opiate receptors by high doses of fentanyl lead to bradycardia in pithed rats. We conclude that in the rat, fentanyl administered intravenously can act at three different levels on cardiovascular regulation: the vagal nerve endings, the brain, and the heart.

Central and peripheral sites for cardiovascular actions of dynorphin-(1-13) in rats.

Dynorphin-(1-13) (i.v.) induced bradycardia and hypotension in artificially ventilated anaesthetized rats. These effects were prevented by MRZ 2266 BS. The bradycardia was inhibited by bilateral vagotomy whereas the fall in blood pressure was not sensitive to bilateral vagotomy and chlorpheniramine treatment. In pithed rats, dynorphin-(1-13) reduced heart rate. This bradycardia was prevented by MRZ 2266 BS but not by tertatolol and cimetidine. It is suggested that the dynorphin-(1-13)-induced effects result from the stimulation of central and cardiac kappa-opiate receptors.

Vagally mediated reflex and cardiac slowing induced by loperamide in rats.

The intravenous injection of loperamide induced an immediate fall in blood pressure and heart rate in anaesthetized rats. Both effects were inhibited by the opiate antagonists naloxone and MRZ 2266 BS. Bilateral vagotomy also inhibited both effects whereas atropine only reduced the bradycardia, but the combination of atropine and tertatolol suppressed the bradycardia. A high dose of loperamide induced bradycardia in pithed rats. This effect was prevented by MRZ 2266 BS but not by naloxone. It is concluded that loperamide can elicit a vagally mediated reflex involving vagal and sympathetic mechanisms and could stimulate cardiac opiate receptors, probably kappa, both effects leading to bradycardia.

Cardiac slowing induced by peripheral kappa-opiate receptor stimulation in rats.

The cardiovascular effects of ethylketocyclazocine were studied in rats anaesthetized with pentobarbital and artificially ventilated. Ethylketocyclazocine (1 mg X kg-1 i.v.) induced a fall in heart rate and blood pressure. The bradycardia and the hypotension were not altered by bilateral vagotomy and atropine, but were inhibited by naloxone and Mr 2266 BS. Ethylketocyclazocine always induced bradycardia in beta-adrenoreceptor-blocked and pithed animals. This bradycardia was prevented by Mr 2266 BS. These results would suggest that a direct peripheral action may occur at kappa-sites, located on the heart after intravenous injection of ethylketocyclazocine in the rat.