Pharmacological characterization of muscarinic receptor subtypes in rabbit isolated tissue preparations.

1 The affinity of some muscarinic antagonists for muscarinic receptors was determined in functional isolated tissue studies in order to compare the muscarinic receptor subtypes in the rabbit. 2 Our attention was specially focused on the question of whether the muscarinic receptors mediating vasodilatation in the aorta resemble or not the ones present on the jejunum of the gastrointestinal tract. 3 Isolated aorta, jejunum, stimulated left atrium and vas deferens preparations of rabbit were investigated with the following muscarinic antagonists: atropine, pirenzepine, methoctramine (N,N'-bis[6-92-methoxybenzyl)amino hexyl]-1,8-octane-diamine tetrahydrochloride) and 4-DAMP (4-diphenylacetoxy-N-methylpiperidine methiodide). 4 The results demonstrate that the receptors on aorta are unlike those on the other rabbit tissues: pirenzepine pA2 was 6.4 on aorta but 8.1 on vas deferens; methoctramine pA2 was 5.9 on aorta but 7.1 on heart; 4-DAMP pA2 was 8.7 on aorta and 8.0 on jejunum. This raises the question: what subtype might be involved?

Cholinergic compounds. VIII Synthesis and biological activity of epiallo-desethermuscarine.

The synthesis of epiallo-desethermuscarine (I) is reported. Its pharmacological activity has been tested and compared with the other carbocyclic analogs of muscarine and its derivatives.

Cholinergic compounds. III - Synthesis and biological activity of epi-desmethyldesethermuscarine, desmethyldesethermuscarine and desmethyldesethermuscarone.

As a further contribution to better understanding of the nature of the cholinergic receptors, desmethyldesethermuscarone (I), epi-desmethyldesethermuscarine (II a) and desmethyldesethermuscarine (II b) were synthesized. With few exceptions, the drop in muscarinic activity of these compounds compared with that of desethermuscarone and desethermuscarines emphasizes the importance of C-7 methyl (5). As far as nicotinic activity is concerned, C-7 methyl appears much less critical.

Cholinergic compounds. V - New synthesis and further pharmacological evaluation of desethermuscarine.

Two new methods of syntesis of (+/-)-desethermuscarine (I) are reported along with the pharmacological evaluation of (I) extended to several muscarinic and nicotinic receptors.

Local anaesthetic effects on trophoblast membrane fluidity.

Previous studies showed that anaesthesia with the barbiturate Thiopental induces an increase in membrane fluidity and a decrease in acetylcholinesterase activity in syncytiotrophoblast plasma membranes (SPM) obtained from placentas after Cesarean section. The aim of the present work was to compare the effect of a local anaesthetic (bupivacaine hydrochloride, trade name Marcaine) on SPM in vivo and to establish whether the anaesthetic is still present in the membrane after tissue preparation. The acetylcholinesterase activity was lower in Marcaine-anaesthetized SPM (27 +/- 3 against 39 +/- 6 in the control). The Marcaine action on the SPM can be ascribed to a competitive inhibition, similar to that reported for Thiopental. Fluorescence studies of the order parameter P showed it to be higher in SPM obtained from control (0.253 +/- 0.012) than in SPM obtained from Marcaine-exposed membranes (0.240 +/- 0.015). The local anaesthetic is still present in the SPM after their preparation (20.1 ng per mg membrane protein). It appears that the local anaesthetic exhibits an effect similar to that of the general anaesthetic, apparently due to binding to the membrane.

Some approaches to the pharmacology of multisubstrate enzyme systems.

Analytical and exploratory in vitro, in situ and in vivo, physio-pharmacotoxicology, from enzymology to population epidemiology, now embraces those approaches that correlate complex dynamic multisubstrate kinetics through conventional and more recent non-invasive quantitative methodologies. Basically, substrates may be classed as pertaining to fundamental energy turnovers (first-order cellular metabolic pathways or networks) and to iso- vs allosteric modulator systems (second-order metabolic control network). Pairs of substrates and cofactors set-up the third-order multienzyme-receptor patterns, which in intact, native in vivo structures establish and maintain the compartmentalized, dynamically superimposed overall coordination of local redox and phosphate potentials. Perturbations of the various levels of the metabolic hierarchy induced by drugs, as well their relaxations, can be readily submitted to non-invasive kinetic analysis. Both indirect and direct titrations of substrate levels, their modelling and statistical ad hoc evaluations of their interrelations can lead to the identification of the multiple sites involved in drug effects as structured at the different orders/levels of concomitant functional variations. Fractal geometries contribute towards defining the space- and time-related events.

Inhibition of the NA(+)/H(+) exchanger reduces rat hepatic stellate cell activity and liver fibrosis: an in vitro and in vivo study.

BACKGROUND & AIMS: The Na(+)/H(+) exchanger is the main intracellular pH (pH(i)) regulator in hepatic stellate cells (HSCs) and plays a key role in regulating proliferation and gene expression. We evaluated the effect of specific inhibition of this exchanger on HSC proliferation and collagen synthesis in vivo and in vitro. METHODS: Rat HSCs were incubated in the presence of platelet-derived growth factor (PDGF), transforming growth factor (TGF)-beta1, iron ascorbate (FeAsc), and ferric nitrilotriacetate solution (FeNTA) with or without the Na(+)/H(+) exchanger inhibitor 5-N-ethyl-N-isopropyl-amiloride (EIPA). pH(i) and Na(+)/H(+) exchanger activity, cell proliferation, and type I collagen accumulation were measured by using the fluorescent dye 2',7'-bis-(carboxyethyl)-5(6)-carboxyfluorescein, by immunohistochemistry for bromodeoxyuridine, and by enzyme-linked immunosorbent assay, respectively. In vivo liver fibrosis was induced by dimethylnitrosamine administration and bile duct ligation (BDL) in rats treated or not treated with amiloride. RESULTS: PDGF, FeAsc, and FeNTA increased Na(+)/H(+) exchange activity and induced HSC proliferation. TGF-beta1 had no effect on the Na(+)/H(+) exchanger and was able, as for FeAsc and FeNTA, to induce type I collagen accumulation. EIPA inhibited all the effects determined by PDGF, FeAsc, and FeNTA and had no effect on TGF-beta1-induced collagen accumulation. In vivo, amiloride reduced HSC proliferation, activation, collagen deposition, and collagen synthesis. CONCLUSIONS: The Na(+)/H(+) exchanger can play a key role in the development of liver fibrosis and in HSC activation in vivo.

Cholinergic compounds VII - Synthesis and pharmacological activity of some trimethylammonium iodides related to desethermuscarine.

Some substances similar to desethermuscarine were synthesized and studied as cholinergics on isolated organs. The results show the importance of the cyclopentane nucleus with regard to activity. The activity of 3-methyltrimethylammonium hexane iodide is particularly interesting for, though lacking the oxygenated function in 3 and having a different methyl spatial arrangement, this compound is only ten times less active than desethermuscarone.