2000 mg/kg altered FOB parameters. CFA at 500 mg/kg altered QTand QTc interval. CFA showed insignificant ulcer score at 500 mg/kg.In conclusion, Celecoxib amorphous formulation did not raised anyconcern related to CNS and GIT safety except prolongation of QTinterval.

doi:10.1016/j.vascn.2009.04.051

In vitro pharmacological profiles of kinase inhibitors: Comparisonwith other pharmaceuticals

Richard W.A. Luke⁎, Mike G. Rolf, Rob H. Bradbury, Ed J. Griffen,Neil J. Hales, Jason Kettle, Andrew P. Thomas, Joanne Bowes,Jean-Pierre Valentin, Andy J. BarkerAstraZeneca UK Ltd, Macclesfield, United Kingdom

Pharmacological profiling is a method for assessing the selectivityof compounds using in vitro assays. It is desirable in drug discovery tominimise the incidence of unwanted toxicities due to off-targetactivities, to reduce the risk of attrition. Kinase inhibitors as a classsuffer from negative perceptions with respect to their safety profiles.In part this is due to the potential for kinase inhibitors to inhibitmultiple kinases in the human genome, not all of which are fullycharacterised or understood. There is also a perception that kinaseinhibitors are generally promiscuous compounds (inhibiting multipleother non-kinase targets). The objective of this study was toinvestigate whether kinase inhibitors are more likely than otherclasses of pharmaceuticals, to interact with non-kinase targets.

The dataset consisted of 758 AZ compounds tested in a set of 591in vitro radioligand binding and enzyme assays, giving 60129datapoints in total. The assays were selected to give broad coverageof target biological space. Compounds were classified as kinaseinhibitors based on the criteria of having an IC50<100 µM in a kinaseassay. Overall promiscuity and hit rates (defined as 3 50% inhibition)in individual assays were compared between kinase inhibitors andother compounds.

199 compounds were classified as kinase inhibitors. Overall hitrates over all compound concentrations were similar between kinaseinhibitors (10%) and non-kinase-targeted compounds (13%); thisresult was not markedly changed if results were limited tocompounds tested at 10 µM, the most common test concentration.Hit rate comparisons for individual assays identified 3 assays wherekinase inhibitors showed significantly higher hit rates than non-kinase compounds at one or more concentrations: adenosinetransporter, adenosine A2A and phosphodiesterase 4 (PDE4).

Data from this analysis demonstrates that compounds that inhibitthe activity of kinases are not inherently more promiscuous thanother classes of compound. A small set of non-kinase targets wasidentified where kinase inhibitors have an increased risk of interac-tion. To avoid toxicities associated with activity at these targets,pharmacological profiling in early drug discovery should be used toidentify and screen out these liabilities.

doi:10.1016/j.vascn.2009.04.052

Role of hyperbaric oxygen therapy on survival and engraftment ofstem cells in the infarct heart

Mahmood Khan⁎, Periannan Kuppusamy, Robert L. HamlinThe Ohio State University, Columbus, OH, United States

Amajor limitation in the applicationof stemcell therapy for ischemicheart disease is the low survival of transplanted cells in the ischemic

hearts. Most of the implanted cells are lost or die within 4 days aftertransplantation into the ischemic heart. Recent studies have shown thatpharmacological preconditioning with hyperbaric oxygen therapy(HBOT) attenuatesmyocardial injuryand also improves cardiac functionin patients with acute myocardial infarction. However, the role of HBOTon survival of transplanted stem cells is not yet studied.

Objective: The objective of this study was to investigate the effectof HBOT (100% O2 at 2 ATA) treated rats on survival and engraftmentof transplanted mesenchymal stem cells (MSCs) in the infarcted ratheart.

Methods: Fisher-344 rats were pretreated with HBOT for 3 daysprior and 10 days after the induction of myocardial infarction (MI) bypermanently ligating the left anterior descending coronary artery(LAD). MSCs (0.5×106 cells/100 µl) were labeled with fluorescentiron oxide probe were implanted in the infarct and per-infarct regionsof the heart. Echocardiography was done at baseline and at 2 weeksafter the MSCs transplantation. Electrocardiogram (ECG) measure-ments from Unipolar Thoracic ECG Lead V3 were done to assess thechanges in ventricular depolarization. Magnetic resonance imaging(MRI) was done to track MSCs in the heart. Immunohistochemicalstaining for iron oxide, CD29 (integrin β), α-sm actin, VWF factor VIIIand VEGF were done in cardiac tissues at the end of 2 weeks.

Results: In MSC+HBOT treated groups there was a significantimprovement in the cardiac function, decreased myocardial infarctionand left ventricular wall thinning compared to MI and MI+MSCstreated groups (Fig. 1). Electrocardiography changes indicated theorientation of the mean QRS vector in the frontal plane tended todeviate to the left in MSCs transplanted hearts compared to MI alone.Prussian blue staining showed the retention of MSCs in the infarctand per-infarct regions of the heart. About 80% of Prussian bluestained cells expressed CD29 (MSCs marker). There was increasedvessel and capillary density in MSC+HBOT treated group comparedto MSC alone. Similarly, VEGF expression was significantly enhancedin HBOT treated group.

Conclusions: Hyperbaric oxygen therapy in conjunction withMSCs improves the survival and engraftment of transplanted MSCsand improves the cardiac function by increased angiogenesis andVEGF expression in the infarct and peri-infarct regions of the heart.The deviation of QRS vectors towards the left side appears to indicatereturn towards electrophysiologic normalcy of the formerly infarctedareas located on the left side of the heart.

doi:10.1016/j.vascn.2009.04.053

Effects of isoprenaline on human stem cellderived cardiomyocytes

S. Bryanta,⁎, C. Wylliea, R. Palmera, R. Heala, J. Demmona, S. Nicola,B. Ansonb, D. Rudy-Reilb, J. MabaVivoMedica (UK) Ltd, Kent, United KingdombCellular Dynamics International Inc, Madison, WI, United States

Cardiac myocytes derived from either primary cultures or humanstem cell sources, in conjunction with microelectrode array (MEA)technology provides an integrated system for assessing cardiotoxicityof early stage lead compounds.

Aims: Isoprenaline (Iso) has multiple effects on the cardiac actionpotential. The aim of this study was to use DrugPrint® software tomeasure and compare the multiple effects of Iso on MEA recordedspontaneous field action potentials from human stem cell derived(hsc) and embryonic rat (erc) cardiomyocytes and assess the use ofhsc with DrugPrint software as an in vitro screening system.

Methods: Hsc and erc myocytes were cultured onto MEAs (220/30ir-ITO-gr, MCS, Germany). sfAPs were recorded with a 60-channel

Abstracts / Journal of Pharmacological and Toxicological Methods 60 (2009) 210–258 219