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Multidisciplinary Approaches to Modern Therapeutics: Joining Forces for a Healthier Tomorrow

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Multidisciplinary Approaches to Modern Therapeutics: Joining Forces for a Healthier Tomorrow Available Online at www.jptcp.com www.cjcp.ca/ ABSTRACTS / RÉSUMÉS Multidisciplinary Approaches to Modern Therapeutics: Joining Forces for a Healthier Tomorrow May 24-27, 2011 Montreal, QC Canadian Society of Pharmacology and Therapeutics La Société canadienne de Pharmacologie et de Therapeutique J Popul Ther Clin Pharmacol Vol 18(2):e315-e363; May 22, 2011 e315 © 2011 Canadian Society of Clinical Pharmacology and Therapeutics. All rights reserved. 2011 CSPT ANNUAL CONFERENCE - ABSTRACTS Contents (Presenters are underlined) Oral Presentations May 26, 2011……………………………………………………………………………..e317 Poster Presentations May 25, 2011……………………………………………………………………………...e320 Poster Presentations May 26, 2011………………………………………………………………………………e338 e316 J Popul Ther Clin Pharmacol Vol 18 (2):e315-e363; May 22, 2011 © 2011 Canadian Society of Clinical Pharmacology and Therapeutics. All rights reserved. 2011 CSPT ANNUAL CONFERENCE - ABSTRACTS ORAL PRESENTATIONS 1Department of Physiology and Pharmacology and 2 THURSDAY - MAY 26, 2011 Division of Clinical Pharmacology, Department of Medicine, The University of Western Ontario, London, Ontario, Canada 1 Corresponding Author: [email protected] Effect of human equilibrative nucleoside Conflict of Interest: None declared transporter 1 (hENT1) expression on adenosine production from neurons Background: The HMG-CoA reductase inhibitors, or Chu S, Parkinson FE statins, are widely prescribed to reduce cardiovascular Department of Pharmacology and Therapeutics, disease risk. There is considerable interindividual University of Manitoba, Winnipeg, Canada variation in statin exposure and response arising from Corresponding Author: [email protected] variability in both transport and metabolism. Conflict of Interest: None declared Objective: Our aim was to better understand the in vivo relevance of the organic anion-transporting Background: Adenosine is produced in brain under polypeptide (OATP) 1B family to atorvastatin (ATV), ischemic conditions. Intracellular and extracellular rosuvastatin (RSV) and simvastatin (SVA) disposition pathways for adenosine formation have been described. in Oatp1b2-/- mice, and the role of metabolism vs Transgenic (Tg) mice with neuron-specific expression transport by comparing ATV, RSV and SVA of hENT1 were developed (Parkinson et al., 2009 J. pharmacokinetics in healthy human subjects given all Neurochem. 109:562-572) to examine neuronal uptake three statins. and release of adenosine during ischemic conditions. Methods: Male Oatp1b2-/- and wild-type mice were Objectives: The present study examined release of dosed 1 mg/kg ATV, RSV or SVA IV, and liver and adenosine and inosine from primary cultures of cortical plasma concentrations measured 30 min later. Ten neurons from wild type (CD1) and hENT1 Tg mice healthy subjects were administered a single oral dose under basal and excitotoxic conditions. of 20mg ATV, 20mg SVA and 10mg RSV in a cross- Methods: Primary neuronal cultures were incubated over study design. Statin acid concentration in plasma with 3H-adenine to radiolabel intracellular ATP. Cells collected 0, 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, 8 and 10 h post- were then treated for 30 minutes (37 ºC) with buffer or dose was measured by LC-MS/MS. N-methyl-D-aspartate (NMDA; 100 µM). The effects Results: Liver-to-plasma ratios were significantly of dipyridamole (DPR; 30 µM), an inhibitor of ENT1 lower in Oatp1b2-/- vs. wild-type mice for ATV and ENT2, α,β-methylene ADP (AOPCP; 50 µM), an (p=0.002) and RSV (p=0.03) but not SVA. In humans, inhibitor of ecto 5’-nucleotidase, and S-(4- plasma exposure of ATV and SVA acid were nitrobenzyl)-6-thioinosine (NBMPR; 100 nM), a significantly related (p<0.05), while RSV profile was selective ENT1 inhibitor, on adenosine and inosine not predictive of ATV or SVA acid exposure. production were assessed. Conclusions: In mice, Oatp1b2 appears important for Results: NMDA significantly increased levels of both the hepatic uptake of ATV and RSV. In humans, ATV adenosine and inosine (p < 0.05); levels were and SVA, which are subject to CYP3A metabolism and significantly greater using hENT1 Tg neurons than transport, appear to share common mechanisms of using CD1 neurons (p < 0.05). DPR, but not AOPCP or elimination, in contrast to RSV, which is not NBMPR, significantly inhibited levels of both significantly metabolised but a substrate of multiple adenosine and inosine (p < 0.05). hepatic uptake and efflux transporters. Conclusions: Intracellular formation and transporter- mediated release of both adenosine and inosine occurs 3 from neurons under ischemia-like conditions. This Validation of the novel in vitro platelet toxicity contrasts with findings from hippocampal slices treated assay (iPTA) for the diagnosis of drug with ischemic conditions, which exhibited transporter- hypersensitivity reactions mediated uptake of adenosine. Elzagallaai AA1,2, Rieder MJ2,3, Koren G1,2,3 1Ivey Chair in Molecular Toxicology, Schulich School 2 of Medicine and Dentistry, University of Western Disposition of atorvastatin, rosuvastatin and Ontario, London, Ontario, Canada; 2Department of -/- simvastatin in Oatp1b2 mice and Physiology and Pharmacology, Schulich School of intraindividual variability in human subjects Medicine and Dentistry, University of Western DeGorter MK1, Urquhart BL1, Tirona RG1,2, Kim Ontario, London, Ontario, Canada; 3Department of RB1,2 Pediatrics, Schulich School of Medicine and Dentistry, J Popul Ther Clin Pharmacol Vol 18 (2):e315-e363; May 22, 2011 e317 © 2011 Canadian Society of Clinical Pharmacology and Therapeutics. All rights reserved. 2011 CSPT ANNUAL CONFERENCE - ABSTRACTS University of Western Ontario, London, Ontario, factor 10a inhibitor, rivaroxaban, was recently Canada. approved for thromboembolism prophylaxis following Corresponding Author: [email protected] total knee or hip replacement. However, rivaroxaban Conflict of Interest: None declared renal elimination is greater than glomerular filtration rate; thus, we hypothesized that the efflux drug Background: Drug hypersensitivity reactions (DHRs) transporter, P-glycoprotein (MDR1), is involved in are rare but potentially fatal adverse events which rivaroxaban excretion and disposition. occur in susceptible patients. The diagnosis and Methods: The ability of MDR1 to mediate rivaroxaban prediction of DHRs is difficult due to their variable transport in vitro was assessed in LL-CPK cells clinical presentation and the overlap of symptoms with overexpressing MDR1 (LMDR1). To determine the in other clinical conditions. Systematic rechallenge, the vivo relevance of MDR1 to rivaroxaban disposition, gold standard for DHRs diagnosis, is not always ethical plasma and tissue concentrations were determined in to perform due to possible serious reactions. Current in Mdr1a deficient mice (Mdr1adef) following oral vitro tests including the lymphocyte toxicity assay administration. (LTA) are cumbersome. We have recently developed a Results: A markedly higher vectorial transport of novel in vitro diagnostic test, the in vitro platelet rivaroxaban was observed in the basolateral to apical toxicity assay (iPTA) for DHRs. direction (B-A) compared to A-B in LMDR1 cells Objective: To validate the iPTA as a diagnostic test for (efflux ratio 5.6, n=5). Additionally, a selective DHRs. inhibitor of MDR1 (LY335979) abolished the B-A Methods: Twenty-eight individuals (14 DHS-sulfa transport of rivaroxaban (efflux ratio 1.2, n=3). patients and 14 healthy controls) were recruited. Blood Following oral administration of rivaroxaban in vivo, samples were obtained and both LTA and iPTA were plasma concentrations did not significantly differ performed independently. Results were then compared between wild-type and Mdr1adef mice (n=6). Liver to to determine the degree of agreement between the two plasma ratio of rivaroxaban concentration was diagnostic approaches. significantly lower in Mdr1adef mice (P<0.01), while Results: There was concentration-dependent toxicity in kidney to plasma ratio was marginally higher compared the cells of patients when incubated with the reactive to wild-type mice. Importantly, rivaroxaban brain hydroxylamine metabolite of sulfamethoxazole for concentrations did not differ, suggesting that other both the LTA and iPTA (p<0.05) and toxicity was efflux transporters at the level of blood-brain barrier significantly greater for the cells of patients versus may be compensating for the absence of MDR1. controls (p<0.05). The two tests had a high degree of Conclusions: Overall, rivaroxaban appears to be a agreement (correlation coefficient: R2 = 0.97). It was substrate MDR1 in vitro. However, further studies are very clear that the iPTA was more sensitive than the required to elucidate additional efflux transporters conventional LTA test in detecting the susceptibility of involved in rivaroxaban disposition in vivo. patient cells to in vitro toxicity. Conclusion: The novel iPTA has considerable 5 potential as an investigative tool for DHS as it is Limited placental transfer of 6-mercaptopurine cheaper to perform and requires no special reagents is mediated by tissue binding and not active that make it more suitable for clinical wider use. transport Hutson JR1,2, Lubetsky A1, Walfisch A1, Garcia- 4 1,2 1,2 Bournissen F , Koren G Role of efflux transporter P-glycoprotein 1Division of Clinical Pharmacology and Toxicology, (MDR1) in rivaroxaban drug disposition 2 1 2 1,2,3 Hospital for Sick Children, Toronto, Insitute of Gong IY , Mansell SE , Kim RB Medical Sciences, University of Toronto,
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