Comparison of the IKr blockers moxifloxacin, dofetilide and E-4031 in five screening models of pro-arrhythmia reveals lack of specificity of isolated cardiomyocytes.

BACKGROUND AND PURPOSE: Drug development requires the testing of new chemical entities for adverse effects. For cardiac safety screening, improved assays are urgently needed. Isolated adult cardiomyocytes (CM) and human embryonic stem cell-derived cardiomyocytes (hESC-CM) could be used to identify pro-arrhythmic compounds. In the present study, five assays were employed to investigate their sensitivity and specificity for evaluating the pro-arrhythmic properties of I(Kr) blockers, using moxifloxacin (safe compound) and dofetilide or E-4031 (unsafe compounds). EXPERIMENTAL APPROACH: Assays included the anaesthetized remodelled chronic complete AV block (CAVB) dog, the anaesthetized methoxamine-sensitized unremodelled rabbit, multi-cellular hESC-CM clusters, isolated CM obtained from CAVB dogs and isolated CM obtained from the normal rabbit. Arrhythmic outcome was defined as Torsade de Pointes (TdP) in the animal models and early afterdepolarizations (EADs) in the cell models. KEY RESULTS: At clinically relevant concentrations (5-12 µM), moxifloxacin was free of pro-arrhythmic properties in all assays with the exception of the isolated CM, in which 10 µM induced EADs in 35% of the CAVB CM and in 23% of the rabbit CM. At supra-therapeutic concentrations (≥100 µM), moxifloxacin was pro-arrhythmic in the isolated rabbit CM (33%), in the hESC-CM clusters (18%), and in the methoxamine rabbit (17%). Dofetilide and E-4031 induced EADs or TdP in all assays (50-83%), and the induction correlated with a significant increase in beat-to-beat variability of repolarization. CONCLUSION AND IMPLICATIONS: Isolated cardiomyocytes lack specificity to discriminate between TdP liability of the I(Kr) blocking drugs moxifloxacin and dofetilide or E4031.

High-resolution functional imaging with ultrasound contrast agents based on RF processing in an in vivo kidney experiment.

Knowledge of the relative tissue perfusion distribution is valuable in the diagnosis of numerous diseases. Techniques for the assessment of the relative perfusion distribution, based on ultrasound (US) contrast agents, have several advantages compared to established nuclear techniques. These are, among others, a better spatial and temporal resolution, the lack of exposure of the patient to ionizing radiation and the relatively low cost. In the present study, US radiofrequency (RF) image sequences are acquired, containing the signal intensity changes associated with the transit of a bolus contrast agent through the microvasculature of a dog kidney. The primary objective is to explore the feasibility of calculating functional images with high spatial resolution. The functional images characterize the transit of the contrast agent bolus and represent distributions of peak time, peak value, transit time, peak area, wash-in rate and wash-out decay constant. For the evaluation of the method, dog experiments were performed under optimized conditions where motion artefacts were minimized and an IA injection of the contrast agent Levovist was employed. It was demonstrated that processing of RF signals obtained with a 3.5-MHz echo system can provide functional images with a high spatial resolution of 2 mm in axial resolution, 2 to 5 mm in lateral resolution and a slice thickness of 2 mm. The functional images expose several known aspects of kidney perfusion, like perfusion heterogeneity of the kidney cortex and a different peripheral cortical perfusion compared to the inner cortex. Based on the findings of the present study, and given the results of complimentary studies, it is likely that the functional images reflect the relative perfusion distribution of the kidney.