Insecticidal arylalkylbenzhydrolpiperidines: novel inhibitors of voltage-sensitive sodium and calcium channels in mammalian brain.

Using preparations derived from whole mouse brain, we have demonstrated that insecticidal arylalkylbenzhydrolpiperidines inhibit the depolarization of synaptoneurosomes as measured by rhodamine 6G fluorescence and block 22Na+ uptake into synaptosomes, when veratridine is used as the activator. These insecticides also have the ability to potently displace the binding of [3H]batrachotoxinin A 20-alpha-benzoate ([3H]BTX-B) to neuronal sodium channels in a concentration-dependent manner. The study compounds can be classified as competitive inhibitors of radioligand binding, since they decrease the affinity of [3H]BTX-B for site 2 without affecting the concentration of sites labelled by this radioligand. Our kinetic analyses revealed that at its IC50, the 4-carbomethoxyaminobenzyl-piperidine analogue reduces the rate of association of [3H]BTX-B with site 2, whereas higher concentrations were required to accelerate dissociation of the [3H]BTX-B:sodium channel complex. These results indicate an ability to interact with both non-activated and persistently activated states of the voltage-sensitive sodium channel, but higher affinity for the former. Such a profile also implies that inhibition of [3H]BTX-B binding to site 2 occurs via an allosteric mechanism. In addition, arylalkylbenzhydrolpiperidines interact with presynaptic voltage-sensitive calcium channels, since we demonstrate that they inhibit increases in [free Ca++] and 45Ca++ uptake when evoked by high KC1 concentration in mouse brain synaptosomal preparations. Such effects generally occur at concentrations that are higher than those required to inhibit sodium channels. Blockade of sodium and calcium channels may therefore contribute to the in vivo neurological effects observed in rodents exposed to these insecticides.

Effects of haloperidol metabolites on neurotransmitter uptake and release: possible role in neurotoxicity and tardive dyskinesia.

This research explored the effects of haloperidol (HP) metabolites on biogenic amine uptake and release, and compared them to those of MPTP and its toxic metabolite, MPP+. In synaptosome preparations from mouse striatum and cortex, the HP metabolites haloperidol pyridinium (HPP+), reduced haloperidol pyridinium (RHPP+), and haloperidol tetrahydropyridine (HPTP) inhibited the presynaptic uptake of dopamine and serotonin, with greater affinity for the serotonin transporter. HPP+ was the most potent inhibitor of dopamine uptake, and HPTP of serotonin uptake, both with IC50 values in the low micromolar range. RHPP+ was less active than the other metabolites, but was more active than the parent compound, HP. Inhibition of uptake was reversed when free drug was removed by centrifugation and then resuspension of the synaptosomes in fresh buffer, suggesting that inhibition of uptake was due to interaction with the transporters and was not due to irreversible cytotoxicity. HPP+ showed noncompetitive inhibition of both serotonin and dopamine uptake, suggesting that it has a relatively slow dissociation rate for its interaction with the transporter proteins. In experiments on amine release, HPP+ and HPTP were four-fold less potent than MPP+ for releasing preloaded dopamine from striatal synaptosomes, and only MPP+-dependent release was antagonized by the uptake blocker, mazindol. In contrast, RHPP+ displayed little ability to release either amine neurotransmitter. HPTP was about two-fold more potent than MPP+ for releasing serotonin from cortical synaptosomes, whereas HPP+ was less active than MPP+. The specific serotonin transport blocker fluoxetine was only able to antagonize release induced by MPP+. These results suggest that HP metabolites bind to the transporters for dopamine and serotonin, but are not transporter substrates. In contrast to their potent effects on amine release, HPP+ and HPTP were unable to release preloaded GABA from cortical synaptosomes. The implications of these results concerning a possible role of HP metabolites in the development of tardive dyskinesia are discussed.