RESUMEN
This study characterises the vasorelaxation and hyperpolarisation effects of the negatively charged quaternary compound tetraphenylboron (TPB) in the rat small mesenteric artery. Segments of rat small mesenteric artery were mounted in a myograph and vessel tone and membrane potential were measured simultaneously. In vessels pre-contracted with vasopressin (0.3-0.6 nM), U46619 (30-90 nM) or methoxamine (0.3-3 microM), TPB (0.1-100 microM) produced a marked endothelium-independent relaxation. However, vasorelaxation responses to TPB were abolished in tissues pre-contracted with K(+) (50 mM), and significantly inhibited by glibenclamide (glib, 10 microM). In the absence of tone, TPB (1-30 microM) caused a concentration-dependent membrane hyperpolarisation of rat mesenteric artery smooth muscle cells, which was not dependent on the endothelium, but sensitive to glibenclamide (10 microM). In methoxamine (0.3-3 microM) pre-contracted vessels, the relaxation response was associated with a marked hyperpolarisation, which was also sensitive to glibenclamide (10 microM), further inhibited by a combination of K(+) channel blockers (glib [10 microM], charybdotoxin [100 nM], apamin [100 nM], 4-aminopyridine [1 mM] and Ba(2+) [30 microM]) and abolished by 50 mM K(+). The results of this study show that TPB causes a vasorelaxation and hyperpolarisation response in the rat small mesenteric artery through a direct action on the vascular smooth muscle. TPB exerts its effects partially via the activation of K(ATP) channels, but also by another mechanism involving K(+)-dependent hyperpolarisation.
Asunto(s)
Arteria Mesentérica Inferior/efectos de los fármacos , Músculo Liso Vascular/efectos de los fármacos , Tetrafenilborato/farmacología , Vasodilatación/efectos de los fármacos , Vasodilatadores/farmacología , Animales , Membrana Celular/fisiología , Relación Dosis-Respuesta a Droga , Técnicas In Vitro , Masculino , Potenciales de la Membrana , Arteria Mesentérica Inferior/fisiología , Músculo Liso Vascular/citología , Músculo Liso Vascular/fisiología , Potasio/fisiología , Bloqueadores de los Canales de Potasio/farmacología , Canales de Potasio/fisiología , Ratas , Ratas Sprague-Dawley , Tetrafenilborato/administración & dosificaciónRESUMEN
Iridium (Ir)-(COD)-pentamidine tetraphenylborate (CAS 225-75-4) was selected from a primary screening to be evaluated in vitro on three Leishmania (L.) strains comparatively to pentamidine used as reference compound. The IC50 values obtained from in vitro evaluation on promastigotes of L. major CRE 26, L. donovani DD8 and L. donovani LV9 were 3.9, 23.5, and 3.3 mumol/l for Ir-(COD)-pentamidine tetraphenylborate and 1.6, 7.7, and 3.9 mumol/l for pentamidine isethionate, respectively. Cytotoxicity on mouse peritoneal macrophages led to determine a chemotherapeutic index of 1.7 for Ir-(COD)-pentamidine tetraphenylborate and 4 for pentamidine. Considering L. donovani DD8, the uptake of iridium complex by the promastigotes was shown to be saturable with a Km value of 17.4 mumol/l and Vmax of 1.3 nmol/mg protein/2 h. After 2 and 4 h incubation of treated promastigotes in drug free medium the absence of Ir-complex efflux is in favour of intracellular drug binding. As a matter of fact iridium complex was shown to bind ribosomal subunits in vitro, with no effect on macromolecular biosynthesis.
Asunto(s)
Iridio , Leishmania donovani/efectos de los fármacos , Leishmania donovani/metabolismo , Compuestos Organometálicos/farmacología , Pentamidina/análogos & derivados , Pentamidina/farmacología , Tetrafenilborato/análogos & derivados , Tripanocidas/farmacología , Animales , Cricetinae , Semivida , Concentración 50 Inhibidora , Leishmania donovani/crecimiento & desarrollo , Leishmania major/efectos de los fármacos , Leishmania major/crecimiento & desarrollo , Leishmania major/metabolismo , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/parasitología , Masculino , Mesocricetus , Ratones , Compuestos Organometálicos/administración & dosificación , Compuestos Organometálicos/farmacocinética , Pentamidina/administración & dosificación , Pentamidina/farmacocinética , Ribosomas/efectos de los fármacos , Ribosomas/metabolismo , Tetrafenilborato/administración & dosificación , Tetrafenilborato/farmacocinética , Tetrafenilborato/farmacología , Tripanocidas/farmacocinéticaRESUMEN
The action of a series of tetraphenylborate ion (TPB) derivatives on yeast cells was studied by electro-rotation of the pre-treated cells. TPB derivatives in which all four phenyl groups were substituted with fluorine, chlorine or trifluoromethyl were much more toxic than the unsubstituted compound, the effect increasing dramatically with increasing size of substituents. These observations suggest that the toxicity of these hydrophobic ions is determined mainly by their size and possibly also by the chemical inductivity of their substituent groups. The order of the toxicities of these ions was in fair agreement with literature values for their translocation rates across artificial bilayers. Incubation times of 3 h were used as standard, longer incubations (up to 48 h) showed that the number of cells affected by low doses of TPB increased with the logarithm of time after the first hour of incubation. Although measurements of the percentage of cells showing co-field rotation showed that controls were not adversely affected by incubations as long as 9 h, rotation spectra showed that some cells suffer loss of internal conductivity during extended incubations. Decrease of the pH of the incubation medium, or inclusion of high concentrations of NaCl or KCl, potentiated the effects of these hydrophobic ions. The toxicity developed slowly, and the sensitivity of the assay was only very weakly dependent on the cell suspension density.