In Vitro Screening for Seizure Liability Using Microelectrode Array Technology.

Drug-induced seizure liabilities produce significant compound attrition during drug discovery. Currently available in vitro cytotoxicity assays cannot predict all toxicity mechanisms due to the failure of these assays to predict sublethal target-specific electrophysiological liabilities. Identification of seizurogenic and other electrophysiological effects at early stages of the drug development process is important to ensure that safe candidate compounds can be developed while chemical design is taking place, long before these liabilities are discovered in costly preclinical in vivo studies. The development of a high throughput and reliable in vitro assay to screen compounds for seizure liabilities would de-risk compounds significantly earlier in the drug discovery process and with greater dependability. Here we describe a method for screening compounds that utilizes rat cortical neurons plated onto multiwell microelectrode array plates to identify compounds that cause neurophysiological disruptions. Changes in 12 electrophysiological parameters (spike train descriptors) were measured after application of known seizurogenic compounds and the response pattern was mapped relative to negative controls, vehicle control and neurotoxic controls. Twenty chemicals with a variety of therapeutic indications and targets, including GABAA antagonists, glycine receptor antagonists, ion channel blockers, muscarinic agonist, δ-opioid receptor agonist, dopaminergic D2/adrenergic receptor blocker and nonsteroidal anti-inflammatory drugs, were tested to assess this system. Sixteen of the seventeen seizurogenic/neurotoxic compounds tested positive for seizure liability or neurotoxicity, moreover, different endpoint response patterns for firing rate, burst characteristics and synchrony that distinguished the chemicals into groups relating to target and seizurogenic response emerged from the data. The negative and vehicle control compounds had no effect on neural activity. In conclusion, the multiwell microelectrode array platform using cryopreserved rat cortical neurons is a highly effective high throughput method for reliably screening seizure liabilities within an early de-risking drug development paradigm.

Chaotic capture of vortices by a moving body. II. Bound pair model.

Previously, we have presented a simple model for the interaction of a fluid vortex structure with a moving bluff body, and demonstrated the existence of a trapping mechanism related to chaotic scattering. This single point vortex model required explicit perturbation to generate chaos and the subsequent complex dynamics. Here, we present a model which attempts to introduce internal degrees-of-freedom in the vortex structure in the simplest manner, by replacing the single vortex with a like-signed pair. We show that this model exhibits chaotic trapping without the need of explicit perturbation, however, the region of parameter space for which trapping occurs is exceedingly small due to the spatially dependent form of the perturbation. We claim that this result explains some the behavior observed in Navier-Stokes simulations of the same vortex-body system, where we find close correspondence between the dynamics of an extended vorticity distribution and the single vortex model. Finally, we generalize the model to unequal strength vortex pairs, and find more complex behavior which includes "partial" capture of the weaker vortex by the body. (c) 1994 American Institute of Physics.