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

Available Online at www.jptcp.com www.cjcp.ca/

ABSTRACTS / RÉSUMÉS

May 24-27, 2011 Montreal, QC

Canadian Society of Pharmacology and Therapeutics La Société canadienne de Pharmacologie et de Therapeutique

Contents (Presenters are underlined)

Oral Presentations May 26, 2011……………………………………………………………………………..e317

Poster Presentations May 25, 2011……………………………………………………………………………...e320

Poster Presentations May 26, 2011………………………………………………………………………………e338

ORALPRESENTATIONS 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,

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, Toronto 1 Department of Physiology and Pharmacology, Corresponding Author: [email protected] 2 Division of Clinical Pharmacology, Department of Conflict of Interest: None for all authors Medicine, 3Lawson Health Research Institute, University of Western Ontario, London, Ontario, Introduction: The immunosuppressant azathioprine is Canada increasingly used in pregnancy for the treatment of Corresponding Author: [email protected] autoimmune diseases or for organ transplant patients. Conflict of Interest: None declared Several studies have demonstrated that azathioprine does not increase the risk for major fetal Background/Objective: Thromboembolic events malformations. Azathioprine is rapidly metabolized to resulting from blood clotting disorders are a significant 6-mercaptopurine (6-MP) and the placenta is source of mortality and morbidity. The novel direct considered a relative barrier to 6-MP. Because 6-MP e318 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 interferes with DNA synthesis, it is important to However, the neonatal safety of oxycodone during determine how the placenta restricts transfer in order to breastfeeding is unknown. identify factors that could increase fetal exposure. Objective: To quantify the incidence of neonatal CNS Objective: To determine if active drug transporters, depression in oxycodone-medicated mothers compared tissue binding, or placental metabolism restrict 6-MP to codeine, and acetaminophen-only group. transfer. Methods: A retrospective study consisting of 3 cohorts Methods: Dual perfusion of a single human placental in 533 breastfeeding mother-infant pairs exposed to lobule ex vivo was utilized and 6-MP was added to the oxycodone (n=139), codeine (n=210) or maternal circulation to determine transplacental acetaminophen-only (n=184) was conducted. A kinetics. 6-MP was also introduced under equilibrative standardized telephone questionnaire was administered conditions to determine if 6-MP is actively effluxed to elucidate adverse maternal and neonatal events into the maternal circulation. Metabolite formation was temporally related to either drug according to maternal also measured during all perfusions. self-reports. Results: At a clinically relevant concentration Results: The incidence of neonatal CNS depression for (50ng/ml), 6-MP appeared in the fetal circulation after oxycodone was 20.1% (28/139) compared to 16.7% 30 minutes. After 180 minutes, the fetal:maternal 6-MP (35/210) for codeine [p>0.05, OR 0.79 95% CI 0.46- concentration ratio was 0.45±0.09 (n=3). After adding 1.38] and 0.5% for acetaminophen (1/184) [p<0.0001, 6-MP to both the maternal and fetal circulations, the OR 46.16 95% CI 6.2-344.2]. Mothers of symptomatic fetal:maternal concentration ratio was 1.205±0.177 neonates in the oxycodone and codeine cohorts took (n=4) after 180 minutes. 6-methylmercaptopurine was significantly higher doses of medication compared to the only metabolite detected and only in perfusions mothers of asymptomatic infants in the same cohorts with 10-fold higher 6-MP concentrations. [oxycodone p=0.0005 (median 0.4 (0.03-4.06) vs. Conclusions: Tissue binding to the placenta, together median 0.15 (0.02-2.25) mg/kg/day and codeine with a short half-life, limits placental transfer of 6-MP. p<0.001 median 1.4 (0.7-10.5) vs. 0.9 (0.18-5.8) Active transport does not play a major role, thus mg/kg/day]. There was significant concordance polymorphisms or drug interactions involving drug between neonatal and maternal CNS depression in both transport proteins are unlikely to leave a fetus more oxyodone and codeine groups [p=0.0006: OR 8.86, vulnerable to 6-MP exposure. 95% CI 2.00-39.24, p<0.001: OR 21.1 95% CI 7.4-60.6 respectively]. 6 Conclusion: Oxycodone is not a safer alternative than Incidence of central nervous system (CNS) codeine in breastfed infants. depression of neonates breastfed by mothers receiving oxycodone for postpartum analgesia 7 Lam J1,2, Kelly L3, Ciszkowski C3,4, Carleton Effects of chronic renal failure on brain drug 5,6 5,7 2 1,2,3,4 transporters in rats BC , Hayden MR , Madadi P , Koren G 1,2 1,2 1 1 Naud J , Michaud J , Beauchemin S , Lefrancois Department of Pharmacology and Toxicology, 1 1 1,2 University of Toronto, Toronto, Canada; 2Division of S , Leblond FA ,Pichette V 1 Clinical Pharmacology and Toxicology, Hospital for Centre de recherche de l’Hôpital Maisonneuve- 2 Sick Children, Toronto, Canada; 3Department of Rosemont, Montréal, Québec; Département de Physiology and Pharmacology, 4Ivey Chair in pharmacologie, Faculté de Médecine, Université de Molecular Toxicology, Schulich School of Medicine Montréal, Montréal, Québec and Dentistry, University of Western Ontario, London, Corresponding Author: [email protected] Canada; 5Child and Family Research Institute, Conflict of Interest: None declared Children’s and Women’s Health Centre of British Columbia; 6Pharmaceutical Outcomes Programme, Background: Many studies demonstrated that chronic Children’s and Women’s Health Center of British renal failure (CRF) significantly affects the expression Columbia, Vancouver, Canada; 7Department of and activity of intestinal, hepatic and renal drug Medical Genetics, Center for Molecular Medicine and transporters. These drug transporters are also expressed Therapeutics, University of British Columbia, in brain cells and at the blood-brain-barrier (BBB) Vancouver, Canada where they limit the entry and distribution of drugs in Corresponding Author: [email protected] the brain. Perturbations in brain drug transporters Conflict of Interest: None declared equilibrium caused by CRF could lead to central toxicity of drugs. Background: Oxycodone has recently replaced codeine for postpartum pain relief in some institutions.

Objective: To evaluate how CRF affects the Objective: To determine the relationship between expression and activity of drug transporters at the BBB various markers of fetal neuropathology and organ- and in the brain using nephrectomised rats. system injury and FAEEs content in meconium of fetal Method: Protein and mRNA expression of influx sheep exposed to ethanol in late gestation. transporters (organic-anion-transporting-polypeptide Methods: From 95-133 days of gestational age (DGA; [Oatp], organic-anion-transporter [Oat]), and efflux term~147 days), chronically catheterized pregnant transporters (p-glycoprotein [P-gp], multidrug- ewes received daily 1-hr infusions of either 0.75 g resistance-related-protein [MRP]) was measured in ethanol/kg (n=13) or saline (n=9). On 134 DGA, ewes CRF and control rat brain biopsies. Intra-cerebral and fetuses were euthanized and fetal tissues, as well as accumulation of radio-labelled benzylpenicillin meconium, were collected for analysis. Meconium (substrate of Oats and MRPs) and digoxine (Oatps, P- FAEEs (palmitic, linoleic, oleic, and stearic) were gp) was used to evaluate BBB permeability to drugs. quantified using headspace solid-phase microextraction Protein expression of transporters was evaluated in rat and gas chromatography-mass spectrometry. brain endothelial cells (RBEC) and astrocytes Results: Total FAEEs content was significantly higher incubated with control and CRF rat serum. in ethanol-exposed fetuses as compared with controls Results: We demonstrated significant 30-50% (P<0.05), with mean content in meconium of 0.174 decreases in protein and mRNA of MRP2-4, Oat3, nmol/g (range 0-0.788) for ethanol and 0.013 nmol/g Oatp2-3 and P-gp in CRF rat brain biopsies, astrocytes (range 0-0.068) for controls. Ethyl stearate and ethyl and RBEC. MRP5 was unchanged. We found a 30% palmitate were measurable only in ethanol-exposed decrease in BBB permeability of benzylpenicillin and fetuses; ethyl oleate was measurable in both ethanol no change in digoxine permeability. We hypothesize and controls, but in higher amounts in ethanol-exposed that similar reductions in the expression and activity of fetuses; while ethyl linoleate was undetectable in both influx and efflux transporters prevented drug groups. accumulation in the brain and that competition with Conclusions: Ethanol exposure in late gestation accumulating endogenous organic anions explains the resulted in elevated FAEEs in meconium of fetal sheep. reduced permeability of benzylpenicillin. Correlational analysis of meconium FAEEs content Conclusion: Even with decreased drug transporters, with post-mortem measures of neuropathology in BBB integrity seems to be conserved in CRF. immersion-fixed fetal brain slices and with fetal organ- system pathology will be conducted to determine 8 whether increased FAEEs content is predictive of Repeated ethanol exposure of fetal sheep in late particular manifestations of ethanol teratogenicity. gestation and fatty acid ethyl esters in meconium Zelner I1,2, Kenna K3, Brien J4, Bocking A5, Harding R6, Walker D3, Koren G1,2 POSTERPRESENTATIONS 1Division of Clinical Pharmacology & Toxicology, The WEDNESDAY - MAY 25, 2011 Hospital for Sick Children, Toronto, ON, Canada; 2 Dept. of Pharmacology and Toxicology, University of 9 Toronto, Toronto, ON, Canada; 3Dept. of Physiology, 4 Thiopurine methyltransferase and inosine Monash University, Clayton, VIC, Australia; Dept. of triphosphate pyrophosphohydrolase genotypes Pharmacology and Toxicology, Queen’s University, Kingston, ON, Canada; 5Dept. of Obstetrics and among patients with inflammatory bowel disease treated with azathioprine Gynaecology, University of Toronto, Toronto, ON, 1,2 1 1 Canada; 6Dept. of Anatomy and Developmental Aljudaibi B , Schwarz UI , Urquhart BL , 1 2 2 2 Biology, Monash University, Clayton, VIC, Australia Dresser GK , Gregor JC , Ponich T , Chande N , Corresponding Author: [email protected] Kim RB1 Conflict of Interest: None for all authors 1Division of Clinical Pharmacology & 2Gastroenterology, Department of Medicine, Schulich School of Medicine Background: Meconium fatty acid ethyl esters and Dentistry at the University of Western Ontario, (FAEEs) are established biomarkers of prenatal ethanol London, Ontario, Canada exposure. We hypothesized that meconium FAEEs Corresponding Author: [email protected] content correlates with ethanol-induced fetal Conflict of Interest: None declared pathology, thus validating their use in identifying newborns at-risk for ethanol-induced disabilities. Background: Azathioprine (AZA) is a well established treatment option for patients with e320 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 inflammatory bowel disease (IBD, ulcerative colitis or 10 Crohn’s disease). However, in clinical practice AZA is The effect of patient education on the incidence not always effective, and nearly 20% of patients of CNS depression in neonates of breast discontinue AZA due to adverse events. Functional feeding mothers taking codeine: preliminary polymorphisms of thiopurine methyltransferase analysis: (TPMT) and inosine triphosphate 1 1 2 2 pyrophosphohydrolase (ITPA), two enzymes involved Chaudhry S , Madadi P , Lausman A , Berger H , Koren G1 in thiopurine metabolism, have been previously 1 associated with toxicity. It has been proposed that Division of Clinical Pharmacology and Toxicology, and Motherisk Program; Hospital for Sick Children, known TPMT and ITPA variant carrier status may 2 predict clinical response and toxicity in IBD patients Division of Obstetric and Gynecology, St. Michael’s treated with AZA. Hospital, Toronto, Canada Aims: To determine if TPMT and ITPA genotype is a Corresponding Author: [email protected] predictor of clinical response and adverse effects Conflict of Interest: None declared (myelotoxicity, hepatitis, pancreatitis, diarrhea and myalgia) among patients with inflammatory bowel Background: Previously, we reported CNS depression disease treated with azathioprine. and one fatality in breastfed neonates of codeine- Methods: Patients diagnosed with IBD undergoing prescribed mothers. This has led us to develop AZA therapy or those previously treated with AZA guidelines which are now routinely given to breast were enrolled. Adverse effects and clinical response feeding mothers taking codeine containing analgesia. were evaluated and correlated with TPMT and ITPA Objectives: We undertook this study to evaluate the genotypes. Genotyping of previously described effect of patient education on the incidence of CNS polymorphisms including TPMT 238G>C (*2 allele), depression in neonates of breastfeeding mothers taking TPMT 460G>A and 719A>G (*3A allele), as well as codeine as compared to our previous studies. ITPA 94C>A, was performed by TaqMan Real-time Methods: This prospective cohort study was PCR and polymerase-chain reaction-restriction conducted between December 2009 and January 2011 fragment length polymorphism (PCR-RFLP). The at St. Michael’s Hospital, in Toronto Canada. The TPMT wild-type allele was designated TPMT*1. breastfeeding mothers taking codeine for postpartum Results: A total of 56 patients were enrolled in the pain relief following Caesarian section were educated study, with 38 patients (67.9%) on therapy and 18 by the study coordinator on the mechanism of action of patients (32.1 %) currently off treatment. Among those, codeine, its possible side effects in mothers, and what 16 patients (28.6%) responded to AZA therapy, 12 signs and symptoms should be monitored in neonate, patients (21.4%) partially responded and 15 patients and recommended duration of codeine use (four days). (26.8%) did not respond to therapy. Among 17 patients Mothers also had 24 hour access to the study physician who experienced adverse events, 10 were unable to in case of concern. Telephone follow up was conducted tolerate AZA therapy. Three patients (5.4%) after 7 days. experienced severe myelosupression (WBC< 2.0 or Results: Out of 220 participants recruited and educated neutrophils <1.0). 4 out of 5 heterozygous carriers for only 2 (0.9%) reported adverse drug reactions in their TPMT*3A developed adverse events compared to 13 infants, as compare to 17 (23.6%) out of 72 (P = out of 48 wild-type carriers (80% vs. 27%, P = 0.032). 0.00005). It is twenty fold lower than the previous 2/4 heterozygous carriers for TPMT*3A responded report. Demographic characteristics like maternal age, well to therapy compared to 14/48 of wild-type (50% parity, gestational age and fetal weight were not vs. 29%, P= 0.58). No TPMT*2 was detected. 3/7 different between the two groups. heterozygous patients for ITPA 94C>A developed Conclusions: Patient education regarding the adverse events compared to 14/46 of wild-type patients maximum duration of codeine use and symptoms of (43% vs. 30%, P = 0.67), and 1/7 heterozygous patients possible adverse reactions, including what actions to for ITPA 94C>A were responders to AZA therapy take, significantly decreases the CNS depression in compared to 15/46 of wild-type patients (14% vs. 33%, infants of breast feeding women taking codeine P = 0.66). containing medications. Conclusion: Our result suggests that genotyping for TPMT may help predict clinical response and adverse 11 events among patients initiated on AZA therapy. Hypertonicity-induced expression of human However, there was no correlation between clinical CYP3A in a humanized transgenic mouse effectiveness and adverse events among patients model carrying the ITPA polymorphism. Chuang AI, Yang MD, Dalvi P Harper PA, Ito S

Division of Clinical Pharmacology & Toxicology, Center, 5Clalit Health Services (Southern District), Hospital for Sick Children, Departments of Paediatrics 6Department of Obstetrics and Gynecology, Beer- and Pharmacology, University of Toronto, Physiology Sheva, Israel, 7Epidemiology and Health Services and Experimental Medicine program, Research Evaluation, Faculty of Health Sciences, Ben-Gurion Institute, Hospital for Sick Children, Toronto, Canada University of the Negev, 8The University of Toronto, Corresponding Author: [email protected] Toronto, Canada, and 9BeMORE collaboration (Ben- Conflict of Interest: None declared Gurion Motherisk Obstetric Registry of Exposure collaboration). Background: The Nuclear Factor of Activated T-cells Corresponding Author: [email protected] 5 (NFAT5) mediates hypertonicity-induced human Conflict of Interest: None declared CYP3A expression in vitro through an enhancer sequence located within intron 2 of CYP3A7. In order Objective: Despite high rates of hypertension disorder to characterize this phenomenon in vivo, we used a during pregnancy, the effects of hypertension on humanized transgenic mouse model that contained the perinatal outcome have not been appropriately entire CYP3A4 and CYP3A7 as transgenes, including separated from those of the medications used. We introns, 5’ and 3’ regulatory elements. First, we evaluated the safety of exposure to antihypertensive observed that neither human CYP3A4-3A7, nor known medications during pregnancy, when accounting for Nfat5 target genes were induced by acute intestinal untreated hypertension. hypertonicity (12h) or acute systemic intra-vascular Study Design: A population- based retrospective hypertonicity (6h), suggesting time-dependency of cohort study was performed, comparing all pregnancies Nfat5 activation in vivo. We then employed prolonged of women exposed and not exposed to antihypertensive hypertonic conditions in the mouse model: a) intestinal medications during pregnancy. A computerized hypertonicity (one week of 8% high-salt diet); and b) database of medications dispensed from 1998 to 2008 prolonged dehydration (one week of dehydration by was linked with computerized databases containing cycling 24h water deprivation and 24h water ad-lib maternal and infant hospitalization records from the recovery phases). Stronger Nfat5 protein expression district hospital during the same period. Multiple was observed in the duodenum after intestinal logistic regression models were performed to control hypertonicity, and in the liver and kidney after for confounders. prolonged dehydration. Consequently, Nfat5 target Results: During the study period 100,029 deliveries gene expression was increased, which is consistent occurred; of those, 620 (0.6%) were exposed to at least with hypertonicity-induced Nfat5 activation. Human one antihypertensive medication (methyldopa or CYP3A4 transgene expression also increased in the atenolol) during pregnancy. A higher rate of low birth duodenum after intestinal hypertonicity (4.7±0.9 fold weight newborns (LBW<2500 grams, 24.4% vs. compared to low salt-diet [0% NaCl]: M±SEM; n=6- 10.3%; p<0.001), intrauterine growth restriction 12, p<0.05), and in the liver (10.8±4.0 fold; n=4-14, (IUGR, 5.2% vs. 2.2%; p<0.001) and preterm delivery p<0.05) and kidney (2.5±0.5 fold; n=4-10, p<0.05) (PTD<37 weeks, 24.4% vs. 8.1%; p<0.001) were noted after prolonged dehydration. Furthermore, human among pregnancies of women who were exposed to CYP3A total protein and activity were increased in antihypertensive medications during the third trimester these tissues. Our findings indicate ambient of pregnancy, as compared to women without hypertonicity-induced human CYP3A4 expression in hypertension and not exposed to antihypertensive vivo, suggesting a novel mechanism of human medications. The association between antihypertensive CYP3A4 expression control. medications (in general), methyldopa and atenolol, and LBW, IUGR and PTD remained significant after 12 adjusting for maternal age, ethnicity, smoking, diabetes Chronic hypertension in pregnancy: perinatal mellitus, lack of prenatal care, multiple pregnancy and outcome in women exposed or unexposed to parity (OR=3.7 95%CI:2.9-4.8; OR=4.3 95% CI:3.0- antihypertensive medications 6.3; OR=3.7, 95%CI: 2.9-4.8 respectively). 1 2 3,4,5 Nevertheless, a similar association was noted while Matok I , Orbach H , Gorodischer R , Koren comparing untreated woman with chronic hypertension G1,8,9, Sheiner E6, Wiznitzer A6, Levy A7,9 1 during pregnancy (n=1074) to woman without chronic The Motherisk Program, Department of Clinical hypertension and not exposed to antihypertensive Pharmacology, Hospital for Sick Children, Toronto, 2 medications (n=97,820) (OR=1.7 95%CI: 1.4-2.0, Canada, Faculty of Health Sciences, Ben-Gurion OR=2.1, 95%CI: 1.5-2.9, OR=1.9; 95%CI:1.6-2.3 for University of the Negev, Beer Sheva, Israel, LBW, IUGR and PTD, respectively) 3Departments of pediatrics, Faculty of Health Sciences, 4 Conclusion: Chronic hypertension with or without Ben-Gurion University of the Negev, Soroka Medical treatment during pregnancy is an independent and e322 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 significant risk factor for adverse perinatal outcomes Morgan AD, Kim RB, Tirona RG such as LBW, IUGR and PTD as compared to births of Department of Physiology and Pharmacology; women without chronic hypertension and without Department of Medicine, Division of Clinical exposure to antihypertensive medications. Pharmacology, Lawson Health Research Institute, The University of Western Ontario, London, Ontario, 13 Canada Intracellular mechanism involved in Corresponding Author: [email protected] downregulation of hepatic cytochrome P450 by Conflict of Interest: None declared chronic renal failure and parathyroid hormone Michaud J1,2, Naud J1,2, Beauchemin S1, Leblond Introduction: It is well appreciated that membrane 1 1,2 transporters play important roles in drug disposition, a FA , Pichette V critical determinant of the pharmacological and 1Centre de recherche de l’hôpital Maisonneuve- 2 toxicological profile of all drugs. P-glycoprotein (P- Rosemont, Montréal, Québec, Canada; Département gp), encoded by the ABCB1 gene, is a clinically de pharmacologie, Faculté de Médecine, Université de important and well-characterized efflux transporter Montréal affecting drug absorption, distribution and elimination. Corresponding Author: [email protected] Despite that a number of in vitro assays for P-gp Conflict of Interest: None declared activity are commonly used, the in vivo relevance of data derived from such assays remains unclear due to Background: Chronic renal failure (CRF) is associated both incomplete knowledge of transporter intrinsic with a decrease in drug metabolism, due to a down- clearance and a lack of predictive extrapolation regulation of hepatic cytochrome P450. Previous strategies. Here we provide the theoretical and studies indicated that CRF modifies activity, protein experimental strategy as well as preliminary data for in and mRNA expression of different P450 isoforms in vitro to in vivo prediction of P-gp mediated transport. vivo and in vitro via circulating mediators. Parathyroid Methods: We have selected sitagliptin, an hormone (PTH) was identified as one of them. CRF unmetabolized drug used in Type 2 diabetes as a P-gp and PTH cause an inhibition of P450. The mechanism probe drug. P-gp transport of sitagliptin was examined remains to be defined. in a model of cultured, polarized epithelial cells Objective: The aim of this study was to evaluate the heterologously expressing varying amounts of contribution of different signaling factors like; PXR, transporter. CAR, NF-κB, PKA and PKC. Results: By monitoring sitagliptin transcellular flux using Methods: Four groups of rats were studied CTL, CRF, liquid chromatography – tandem mass spectrometry in CRF with parathyroidectomy (CRF-PTX) and CTL- combination with mathematical modeling, we obtain a PTX. Liver and hepatocyte CAR and PXR mRNA value for P-gp intrinsic clearance. This intrinsic transport expression was measured by qPCR and their protein clearance is normalized to P-gp protein content of cells as expression was measured by Western blots. Cultured determined by quantitative proteomic analysis. hepatocytes were incubated with CTL, CRF, CRF-PTX Conclusion: These data, which would be the first of and CTL-PTX sera, or with or without PTH, and their kind, are expected to form the foundation for nuclear extracts were obtained. Nuclear extracts were quantitative methods for in vitro to in vivo prediction used for NF-κB flux cytometry and Western blots (p50 of drug pharmacokinetics and ultimately therapeutic and p65). efficacy. Results: We observed down-regulations of PXR and CAR protein (43%, 44%, respectively) and mRNA (40%, 42%, respectively) CRF rat’s liver. PTX 15 prevents the mRNA down-regulation of CAR and PXR A comparison of folic acid pharmacokinetics in in CRF rats. We observed a NF-kB accumulation in obese and non-obese women of childbearing liver’s nuclei of CRF rats. Furthermore, blocking NF- age κB with inhibitors counteracts the effect of CRF and Stern SJ1,2, Matok I2, Kapur B1,2,3,4, Koren G1,2 PTH on P450 expression. 1Department of Pharmacology, University of Toronto; Conclusion: In conclusion, CAR, PXR and NF-κB 2Division of Clinical Pharmacology and Toxicology, could be implicated in the down-regulation of hepatic The Hospital for Sick Children; 3Department of P450 by CRF and PTH. Clinical Pathology, Sunnybrook Health Science Centre; 4Department of Laboratory Medicine and 14 Pathobiology, University of Toronto, Toronto, Canada Quantitative in vitro to in vivo prediction of P- Corresponding Author: [email protected] glycoprotein-mediated drug transport Conflict of Interest: None declared

Background: While maternal folate deficiency during Methods: Bare, oleic acid and bovine serum albumin the periconceptional period represents a major risk (BSA) coated IONPs were examined in a mouse brain factor for neural tube defects (NTD), obesity has been microvessel endothelial cell line (bEnd.3) and mouse recognized as an additional risk factor. Studies have primary cultured neurons and astrocyte preparations in identified an increased risk for NTD-affected births the presence and absence of a magnetic field. among obese mothers even after adjusting for folic acid Accumulation of IONPs was examined over a 2 hour supplementation. However, while folic acid intake may period using Prussian blue staining. Cytotoxicity was have been at the recommended dose in these samples, assessed by addition of 3-(4,5-Dimethylthiazol-2-yl)- blood folate concentrations were not monitored to 2,5-diphenyltetrazolium bromide (MTT) after 24-hour ensure protective levels were reached. exposure to various concentrations of formulated Objectives: To compare folic acid pharmacokinetics in IONPs. obese and non-obese women of childbearing age. Results: In neurons, oleic acid coated IONPs had the Methods: Healthy obese (n = 12) and non-obese (n = greatest cell association compared to BSA coated 12) women of childbearing age volunteered to IONPs. In contrast, rank order of cell association for participate. Each obese participant was matched to a IONPs in astrocytes was uncoated > oleic acid > BSA non-obese participant, and assigned an equivalent dose coated. The presence of magnetic field resulted in more per kilogram body weight of folic acid. Folic acid was IONPs within the cells regardless of formulation. None orally administered after a 6-hour fast, and blood of the formulations produced significant (i.e. greater samples were taken over a 10-hour period to evaluate than 10%) toxicity at concentration up to 100ug/mL. pharmacokinetic parameters. Neurons appeared more sensitive to oleic acid coated Results: Area under the curve (AUC) was found to be IONPs at concentrations above100 ug/mL. significantly higher in the obese group (P = 0.008). Conclusions: The various formulations of IONPs were Defining AUC as a function of dose per lean body safe in all cells examined at concentrations less than weight (LBW) was found to be a stronger predictor 100 ug/mL. Accumulation and cytotoxicity of IONPs than dose per total body weight (r2 = 0.90 and 0.76, was increased in the presence of a magnetic field. respectively). Conclusions: This indicates that the body tightly 17 controls systemic exposure to folic acid, with 90% of Down-regulation of human cytochrome P450 variability in AUC controlled by the dose per LBW. 2C8 by 3-methylcholanthrene Periconceptional supplementation recommendations Utgikar R, Riddick DS may need to be adjusted to account for LBW Department of Pharmacology and Toxicology, differences in the obese population. University of Toronto, Toronto, Ontario Corresponding Author: [email protected] 16 Conflict of Interest: None to declare Examination of superparamagnetic iron oxide nanoparticle accumulation and toxicity in Background: 3-Methylcholanthrene (MC) is a model various brain related cell culture models. polycyclic aromatic hydrocarbon that induces Sun Z, Yathindranath V, Hegmann T, Miller DW cytochrome P450 1A1 (CYP1A1) expression via aryl Department of Pharmacology and Therapeutics, hydrocarbon receptor activation. MC also down- Faculty of Medicine, University of Manitoba, regulates certain constitutive hepatic P450s in rodents, Winnipeg, Manitoba, Canada but the mechanism and human relevance of this Corresponding Author: [email protected] response remain poorly understood. Recent reports Conflict of Interest: None declared suggest that MC down-regulates the expression of CYP2C8 in primary human hepatocytes. This has Background: Superparamagnetic iron oxide potential clinical importance because human liver nanoparticles (IONPs) have shown great promise in CYP2C8 metabolizes therapeutic agents, such as the biomedical imaging and drug delivery. However, antineoplastic paclitaxel, and endogenous signalling issues pertaining to the toxicity of IONPs are a concern molecules, such as all-trans-retinoic acid. especially for those applications involving IONPs and Objectives: To determine if MC alters the expression the brain. of CYP2C8 at the mRNA level in two human Objectives: To determine whether cell toxicity was hepatocellular carcinoma cell lines, HepG2 and correlated with cell accumulation of various IONP HepaRG. formulations and to examine the impact of magnetic Methods: Cells were treated with vehicle or MC (1 or field on both these parameters. 5 µM) for 24 or 48 h. Cytotoxicity was assessed by

e324 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 trypan blue dye exclusion. CYP1A1 and CYP2C8 48%, respectively) compared to controls (P < 0.05). mRNA levels were measured by real-time RT-PCR. CYP3A2 protein expression was significantly Results: Under conditions that resulted in minimal decreased in both experimental groups compared to cytotoxicity, MC induced CYP1A1 mRNA levels in controls (P<0.05). Vmax values for midazolam HepG2 cells by approximately 245- to 425-fold. MC at metabolism were weakly correlated with day 42 serum 5 µM suppressed CYP2C8 mRNA levels in HepG2 creatinine levels (P < 0.05). cells by approximately 78% at 24 h and this response Conclusions: Our results demonstrate that CYP3A did not persist at 48 h. Similar studies are in progress activity and expression is decreased in mild renal using HepaRG cells as a more differentiated model that failure, which suggests that drug therapy for patients in maintains higher basal levels of several constitutive early stages of kidney failure may be compromised for P450s. drugs that are substrate for CYP3A. Conclusions: The demonstration of CYP2C8 mRNA suppression by MC in human liver-derived cell lines 19 will facilitate our efforts to define the molecular Mechanism of convolvulus arvensis induced mechanisms and functional impacts of the modulation relaxation of thoracic aorta in rabbit of this important human P450 by environmental Al-Aghawani W1, Al-Madi S2, AL-Lahham A2, toxicants. Abu Jayyab A1 1Faculty of Pharmacy, International University for 18 Science & Technology, Syria; 2Faculty of pharmacy, Hepatic drug metabolism in varying degrees of Damascus University, Damascus, Syria renal function using rat models of renal failure Corresponding Author: [email protected] 1 1 2,3 2,3 Velenosi TJ , Fu A , Luo S , Wang H , Conflict of Interest: None declared Urquhart BL1,3 1Schulich School of Medicine and Dentistry, Background: Convolvulus arvensis L plant (CAL) L is Department of Physiology and Pharmacology, The shown to produce relaxation in some smooth muscle. It University of Western Ontario, London, Ontario, is used in traditional herbal medicine for many Canada; 2Department of Surgery, The University of purposes, especially as anti-spasmodic in Middle East. Western Ontario, London, Ontario, Canada; 3Lawson Objective: Experiments were undertaken to determine Health Research Institute, London, Ontario, Canada whether ethanolic extract obtained from CAL has Corresponding Author: [email protected] vasorelaxant activity in the rabbit aorta rings and, if so, Conflict of Interest: None declared to elucidate the underlying mechanism. Methods: Rabbit aorta rings were suspended in organ Background: Chronic renal failure (CRF) is the result chambers for the measurement of changes in isometric of decreasing renal function over time. Expression and tension in the presence of pheylephrine or activity of drug metabolizing enzymes such as CYP3A glibenclamide. are decreased in end stage renal disease. However, only Results: CAL decreased vessels contraction by a small percentage of patients with CRF are at the final phenylephrine in both presence and absence of an stage of the disease. intact endothelium groups compared with the control Objectives: This study aimed to determine the changes group (IC50 = 358 mg/ L with endothelium and 399 in drug metabolizing enzyme function and expression mg/ L without endothelium, P< 0.05). In addition, in rats with varying degrees of renal failure. pretreatment of aortic rings with glibenclamide Methods: Sprague-Dawley rats underwent either 2/3 inhibited partially CAL induced relaxation in or 5/6 nephrectomy by partial left kidney resection Phenylephrine–contracted aortic tissues. followed by complete right nephrectomy. Control rats Conclusions: These data indicate that calcium might underwent sham laparotomies. Rats were sacrificed on play an important role in the mechanism of CAL day 42 and CYP3A activity was determined in liver induced relaxation of aorta tissues. Also, it might be microsomes by evaluating midazolam metabolism involved, at least in part, potassium channel in the using ultra-performance liquid chromatography with relaxation activity of CAL. The present findings photodiode array detection. suggest that CAL could be a candidate of herbal Results: On day 42, serum creatinine levels were 23.0 medicine for cardiovascular diseases associated with ± 1.4, 37.4 ± 1.0 and 75.4 ± 14.7 M in control, 2/3 aorta tissues dysfunction. and 5/6 nephrectomized rats, respectively. Vmax values for 4-OH and 1-OH midazolam formation were lower 20 in 2/3 nephrectomized rats (29% and 45%, In vitro metabolism study of specific cyp450 respectively) and 5/6 nephrectomized rats (36% and substrates in breast cancer cell lines

Armstrong C1,2, Belanger F2, Gaudette F2, Balicki Background: Interferon alpha (INF) is an effective D3, Turgeon J1,2 treatment for a variety of conditions. Since these 1Faculty of Pharmacy, Université de Montréal, conditions could happen in pregnancy, Information Montreal, QC, Canada; 2CRCHUM, Centre de regarding the safe use of this medication in pregnancy Recherche du Centre Hospitalier de l’Université de is essential. This systematic review attempts to Montréal, QC, Canada; 3Faculty of Medicine, summarize all published data on outcomes of INF Université de Montréal, Montreal, QC, Canada alpha exposed pregnancies. Corresponding Author: Methods: Using key words INF alpha, pregnancy, we [email protected] searched Pub Med, EM BASE, and Google, since INF Conflict of Interest: No conflict of interest to report alpha was introduced in the market. We were able to locate only case reports of INF alpha exposure in Background/Objectives: A treatment failure is pregnancy. All cases were collected and included in commonly observed in breast cancer patients due to an our review. We also collected 71 cases that were innate or acute resistance to anti-cancer agents. diagnosed with essential thrombocythemia, but did not Understanding, the local metabolism of anti-cancer receive any medication in pregnancy. agents by CYP450s in breast cancer cells could help in Results: Among 61 INF alpha exposures in pregnancy the development of a more targeted treatment we located, mean maternal age was 31±4 years, median approach. Our goal is to evaluate if measurable 33 years, and range 23-43 years. Mean full term metabolism of specific CYP450 substrates is present in babies’ weight was 3010, median 2680 grams and the breast cancer cell lines, and to compare this range 1350-3800. Mean gestational age at delivery was metabolism to the expression of CYP450 mRNAs. 37±3 weeks, median 38, range 30-41 weeks. No major Method: Seven commonly used breast cancer cell lines malformation or stillbirth was reported. There was one (and one normal breast cell line) were cultured and spontaneous abortion, and 12 preterm deliveries (20 % then plated in 24 well plates. Once confluent, the cells of all the cases). In 71 cases with the same underlying were incubated in the presence of specific substrates, diseases who did not receive any medication in and the metabolism was determined by measuring the pregnancy, 46 out of 71 had early or late abortion. appearance of metabolites by LC-MS-MS. In order to There were 3 cases of stillbirth and 4 cases experienced correlate metabolism with mRNA expression, RNA preterm delivery. 18 cases had term normal babies. was isolated from each cell line, and the mRNA Conclusion: This systematic review suggests that INF relative expression was determined by RT-PCR alpha does not increase the risk of abortion, major (TaqMan Assay) for 19 CYP450 isoforms. malformation, still birth, and premature deliveries, and Results: RT-PCR results demonstrated that each cell probably it has a protective effect on prevention of line showed differential expression of CYP450 early/late spontaneous abortion and still birth in this mRNAs. The expression of CYP2J2 is of interest particular population. because several cell lines showed elevated expression of this isoform, such as MCF-7 cells, whereas others, 22 such as Hs578T cells, showed little expression. The Motherisk cancer in pregnancy forum: a Metabolism was observable in cells lines for the unique way of counseling specific CYP2J2 substrate, Ebastine, where MCF-7 Carey N1,2, Koren G1,2,3 cells demonstrated measurable metabolism in as little 1Division of Clinical Pharmacology and Toxicology, as 30 minutes. Hospital for Sick Children, Toronto, Canada; Conclusions: Our results suggest that the local 2Department of Pharmacology and Toxicology, metabolism of various anti-cancer agents could be University of Toronto, Toronto, Canada; 3Ivey Chair in significant enough to greatly impact the resistance that Molecular Toxicology, Schulich School of Medicine is observed in breast cancer patients. and Dentistry, University of Western Ontario, London, Canada 21 Corresponding Author: [email protected] A systematic review of the fetal safety of Conflict of Interest: None declared interferon alfa Yazdani Brojeni P, Matok I, Garcia Bournissen F, Background: Cancer occurs in approximately 0.07- Koren G 0.1% of pregnancies. The diagnosis of cancer in Motherisk Program, Hospital for Sick Children, pregnancy complicates optimal treatment due to University of Toronto, Toronto, Canada potential risk to the fetus. The Motherisk program Corresponding Author: [email protected] established The Consortium of Cancer in Pregnancy Conflict of Interest: None declared Evidence (CCoPE) in an effort to address the e326 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 deficiency in information regarding the care of cancer Objective: To describe the frequency and severity of and associated therapies in pregnancy. Subsequently, a ototoxicity in paediatric patients treated with cisplatin Cancer in Pregnancy Forum was created to provide based chemotherapy in a third level paediatric hospital unique, evidence-based counseling to women and their in Mexico City. medical professionals. Methods: Three audiology evaluations were Objectives: To describe the characteristics of the cases prospectively conducted in paediatric patients (<18 counseled by the Cancer in Pregnancy Forum. years) with solid tumour cancers, at baseline and at the Methods: Review of cases documented in the Cancer end of each of two chemotherapy cycles. Audiology in Pregnancy Forum since its inauguration. Data were data was used to classify hearing loss according to the collected on the person who initiated the post, the type Common Terminology Criteria for Adverse Events. of cancer, the treatment plan, response time and Results: Forty-two patients participated in this study, completeness of information. aged 4 to 17 years, 52% were female, being Results: There were a total of 129 inquiries on the osteosarcoma (78.6%) the predominant cancer type. Cancer Forum from 1999 to 2011. Healthcare Bilateral hearing loss was observed in 31% of patients providers and scientists initiated 41% of the postings, at the end of the first chemotherapy cycle, while 62% whilst women or their partners made up the remaining report hearing loss at the end of the second cycle, 59%. Inquiries regarding maternal chemotherapy were mainly in frequencies over 4,000hz (p<0.001). This the most frequent (30%), followed by questions represent an increased risk of hearing loss (OR= 2.00, concerning specific cancers (20%), radiation (17%), [CI 95% 1.26-3.17], p=0.005) from the first to the paternal chemotherapy (13%), other therapies (12%), second cycle of chemotherapy. Cumulative dose, age, breastfeeding (5%), female fertility (1.5%) and male treatment scheme and tumour type were not related to fertility (1.5%). hearing lose. Conclusions: A diagnosis of cancer during pregnancy Conclusion: Hearing loss in Mexican children under is a major stress for the expecting mother and her cisplatin therapy is similar to that reported previously family. The Cancer in Pregnancy forum is the only in other populations. Although ototoxicity was related forum of its kind worldwide, providing women and to chemotherapy cycle number, it cannot be positively medical professionals evidence-based information correlated with cumulative dose of cisplatin, tumour regarding diagnosis, treatment, symptoms and other type or age. Further research is required to correctly concerns with respect to cancer during pregnancy and characterize ototoxicity in Mexican patients. lactation. 24 23 Statins and their category X pregnancy Ototoxicity in Mexican children receiving classification: pravastatin may merit cisplatin based chemotherapy reconsideration for a novel indication 1,2 3 Castelan-Martinez OD , Garcia-Rojas Y , Fierro- Davidovits J1,2,3, Laskin CA2,3,4, Casper RF3,4, 3 3 1 Evans M , Marques-Avila S , Rivas-Ruiz R , Clark CA2,4, Spitzer KA2,4, Koren G1,3 4 2 Jimenez-Mendez R , Castaneda-Hernandez G , 1Hospital for Sick Children; 2LifeQuest Centre for Carleton, B4 Reproductive Medicine; 3University of Toronto; 1Unidad de Epidemiologia Clinica, HIMFG, Mexico; 4Mount Sinai Hospital, Toronto, Canada 2Departamento de Farmacologia CINVESTAV-IPN, Corresponding Author: [email protected] Mexico; 3Servicio de Audiologia y Foniatria, HIMFG, Conflict of Interest: None declared Mexico; 4Department of Paediatrics, University of British Columbia, Canada Background: Animal models of recurrent pregnancy Corresponding Author: [email protected] loss (RPL) and preeclampsia have implicated elevated Conflict of Interest: None declared levels of tissue factor (TF), and resultant hypercoagulability and inflammation, as a key factor in Introduction: Cisplatin has been widely used as a placental and fetal damage and pre-eclampsia. Using a chemotherapeutic agent for a variety of paediatric mouse model that shares features with human RPL and malignancies. One of the most incapacitating adverse pre-eclampsia, treatment with pravastatin (HMG CoA drug reactions reported on patients under cisplatin reductase inhibitor) down-regulated TF and rescued therapy is ototoxicity, associated with permanent pregnancies. However, the FDA has given a category bilateral hearing loss. Even though extensive X classification for all statins (i.e. contraindicated in publications are available in the literature, information pregnancy). Yet, unlike lipophilic statins that comprise on Latin-American patients is scare. the majority of this drug group, pravastatin is

J Popul Ther Clin Pharmacol Vol 18 (2):e315-e363; May 22, 2011 e327 © 2011 Canadian Society of Clinical Pharmacology and Therapeutics. All rights reserved. 2011 CSPT ANNUAL CONFERENCE - ABSTRACTS hydrophilic and hepatospecific, and therefore less encountered with generics. Additional articles were likely to enter fetal circulation. obtained by searching the bibliographies of relevant Objective: To evaluate the FDA category X references. classification for pravastatin. Results: Formulation substitution of Methods: Literature review of teratogenic effects of anticonvulsants/mood stabilizers (carbamazepine, pravastatin exposure. valproate, lamotrigine, gabapentin, topiramate, lithium) Results: Contraindication of statins in pregnancy is antidepressants (amitriptyline, nortriptyline, based on animal testing and a small number of case desipramine, fluoxetine, paroxetine, citalopram, reports involving two lipophilic statins, lovastatin and sertraline, mirtazapine, bupropion, venlafaxine) simvastatin. Increased risk of teratogenicity has not antipsychotics (clozapine, risperidone) and anxiolytics been established. Contrary to lipophilic statins, (clonazepam, alprazolam) has been linked to clinical pravastatin did not exhibit teratogenicity in animal deterioration, decreased tolerability/toxicity and testing or human cases. Pravastatin has not been shown pharmacokinetic changes. Decreased compliance may to be teratogenic in multiple cohort studies. result from a change in formulation. Generic Conclusions: Despite the lack of established substitution is not always economically profitable teratogenic risks, pravastatin remains contraindicated when the consequences of relapses and decreased in pregnancy. Since suspension of compliance are taken into account. hypercholesterolemia therapy during gestation is Conclusions: Publication bias and heterogeneity of the considered safe, this contraindication has not studies are limitations of this review. It is yet previously been questioned. However, promising new premature to determine the true equivalence between indications for prevention of adverse pregnancy generic and original medications. Caution is still outcomes and reassuring reports of its safety during advised when a patient’s medications are switched to gestation support both reevaluation of its another formulation. Health professionals should be contraindication and implementation of controlled sensitized to the possible consequences of generic clinical studies to establish the efficacy of pravastatin substitution. for treatment of RPL and preeclampsia. 26 WITHDRAWN 25 The epidemiology of intentional self-poisoning: Switching from brand-name to generic a population-based study psychotropic medications: a literature review Finkelstein Y, Sivilotti MLA, Lam K, Hutson JR, Desmarais JE, Beauclair L, Margolese HC Koren G, Juurlink DN McGill University Health Centre, Clinical Hospital for Sick Children, University of Toronto; Psychopharmacology and Therapeutics Unit, Montreal, Queen’s University, Kingston; Institute for Clinical Quebec, Canada Evaluative Sciences, Toronto, Canada Corresponding Author: Corresponding Author: [email protected] [email protected] Conflict of Interest: None declared Conflict of Interest: None declared 27 Background: Current world economics encourage use CYP2C19 genotyping in two infants with of generic medications which are less expensive than gastroesophageal reflux brand name originals. There is however controversy as Fujii H1, Sakaguchi S1, Garcia-Bournissen F1, to their clinical equivalence. Clinical deterioration, 2 2 1 adverse effects and toxicity have been described with Gaedigk A , Leeder S , Ito S 1Clinical Pharmacology and Toxicology, The Hospital generic substitution. 2 Objective: To explore issues about generic substitution for Sick Children, Toronto; Clinical Pharmacology & of psychotropic medications reported in the literature. Therapeutics, Children's Mercy Hospital, Kansas City Methods: Pubmed was searched from January 1, 1974 Corresponding Author: [email protected] to March 1, 2010. The MeSH term “generic, drugs” Conflict of Interest: None declared was combined with “anticonvulsants”, “mood stabilizers”, “lithium”, “antidepressants”, “antipsychotics”, Background: Although the impact of CYP2C19 “anxiolytics” and “benzodiazepines.” Articles in English, genotypes on PK of proton pump inhibitors (PPI) has French, or Spanish were considered if they discussed been extensively investigated, clinical utility of clinical equivalence of generic and original CYP2C19 genotyping is still not clear. Also, the medications, generic substitution, or issues about optimal PPI dosage for children under 2 years old has effectiveness, tolerability, compliance, or economics not been yet determined. Here we show 2 infant cases e328 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 with severe gastroesophageal reflux who failed to “transplantation” and “children”. References of respond to high dose of pantropazole. relevant articles were screened as well. Case 1; 11 months old (5.6kg). Pantoprazole was Results: Fifteen of the 87 articles were considered administered via continuous infusion 8.5mg/kg/day relevant. Ten papers researched liver transplant (standard dose: 1mg/kg/day), but gastric pH remained recipients, 4 kidney transplant recipients and one low. CYP2C19 genotyping showed that the patient was abdominal and thoracic transplant recipients. The a heterozygote of CYP2C19 wild type (*1) and high incidence of tacrolimus-induced nephrotoxicity ranged function genotype (*17). Apparent clearance was from 0%-76.5%. This range is a direct result of the 0.37L/kg/h (reported clearance at 2-4 y.o; 0.20±0.23 differences between the studies. Follow-up times L/kg/h). Pantoprazole was then switched to 15mg tid ranged from pre-transplant until 8 years post- oral omeprazole with successful therapeutic response. transplantation. However, five studies did not report Case 2: 5 months old (5.4kg). She received their follow-up times. Seven studies did not clarify 14.0mg/kg/day of pantoprazole without response. Her their definition for tacrolimus nephrotoxicity. Two of CYP2C19 genotype was *1/*1 (wild type). these studies did not mention the type of tacrolimus Pantoprazole was changed to oral omeprazole with toxicity reported. Those studies reporting a definition remarkable therapeutic response. for tacrolimus-induced nephrotoxicity, mainly used Discussion: These 2 infants required 8.5 times and 14 grading systems based on GFR. One study used times higher doses of pantoprazole than normal histological grading systems. However, cut-offs for recommended dose for infants (1mg/kg/day). The high nephrotoxicity were not reported in any of the grading clearance was confirmed in Case 1, which is consistent systems. with CYP2C19*17 genotype. On the other hand, Case Conclusions: Due to the many differences between the 2 suggests a wide range of functionality within the studies, a decisive incidence number cannot be given. CYP2C19 *1/*1 genotype and/or pharmacodynamics Further studies need to address the implications of (PD) variations. these different definitions on the incidence number to Conclusions: CYP2C19*17 genotype may be have a better understanding of the vastness of the sufficient but not necessary as a cause of pantoprazole problem. therapeutic failure in infants. Studies of CYP2C19 genotype - pantoprazole PK/PD correlation are needed 29 to determine the optimal dose for infants. Assessing interactions between herbal medicines and drugs: a review 28 Girard L1, Filippelli A2, Boon H3, Gardiner P4, The incidence of tacrolimus-induced Vohra S1,5 nephrotoxicty in children: do we really know? 1CARE Program, Department of Pediatrics, Faculty of 1,2 2,3 1 Gijsen VMGJ , Koren G , de Wildt SN Medicine, University of Alberta, Edmonton, Alberta; 1Intensive Care and Department of Pediatric Surgery, 2School Of Public Health, Boston University, Boston, Erasmus MC Sophia Children’s Hospital, Rotterdam, Massachusetts; 3Leslie Dan Faculty of Pharmacy, the Netherlands; 2Division of Clinical Pharmacology University of Toronto, Toronto, Ontario; 4Department and Toxicology, the Hospital for Sick Children, of Family Medicine, Boston University Medical Toronto, Canada; 3Ivey Chair in Molecular Toxicology, Center, Boston; 5School of Public Health, Department Department of Medicine, University of Western of Pediatrics, and Department of Medicine, University Ontario, London, Canada of Alberta, Edmonton, Alberta Corresponding Author: [email protected] Corresponding Author: [email protected] Conflict of Interest: None declared Conflict of Interest: None declared

Background: Tacrolimus is a calcineurin-inhibitor and Background: Herbal products are generally considered the drug of choice for pediatric solid organ transplants. safe, but not necessarily when used concurrently with Although the renal effects of calcineurin-inhibitors drugs. A reference tool for rapid identification of have widely been studied, the data on tacrolimus- known herbal medicine-drug interactions was created induced nephrotoxicity in children is limited. to help clinicians. Objectives: To determine the incidence of tacrolimus- Objective: To review the herb-drug literature and induced nephrotoxicity in children by systematically update the herb-drug interaction grid. reviewing the literature. Methods: Searches were conducted in MEDLINE and Methods: Pubmed/Medline, Embase and Google were EMBASE between 2007 and 2010, using the following searched from their inception till February 2nd 2011 search terms: 34 commonly used herbs combined with with the search terms “tacrolimus”, “nephrotoxicity”, ‘clinical trials’, ‘case studies’, and ‘case reports’.

Reference lists of relevant review articles were pro-angiogenic genes was evaluated by real-time pcr. analyzed for additional papers, and a tertiary source Results: We detected a rapid accumulation of was consulted to identify other herb-drug interactions. succinate following the insult. GPR91 was primarily Data extraction included the amount of evidence localized in neurons within the cerebral cortex. To regarding the severity and likelihood of the interactions determine the role of succinate in this injury paradigm, between each herb and each category of we injected succinate intraventricularly in newborn pharmaceutical agents. The interactions were classified animals and detected a 30% increase in blood vessel into four groups: 1) No reported or theoretical formation which was corroborated ex vivo with a robust interactions, 2) Theoretical interactions based on increase in vessel density in cortical explants. animal of in vitro data, 3) Theoretical interactions Stimulation of primary neuronal cultures with succinate extrapolated from clinical data, and 4) Interactions lead to a 2.1-fold increase in VEGF mRNA. supported by clinical evidence. Conclusion: Collectively our data demonstrate that Results: A total of 1553 references were identified by succinate strongly affects the cerebral vascular the searches. 1514 articles have been screened, and 120 response to ischemia. These results offer a new and have been included for extraction. Full results will be potentially important target to optimize recovery ready in October. following cerebral vascular injury. Conclusions: The herbal medicine-drug interaction grid will allow clinicians to have a guide on potential 31 harms based on the most recent literature. This quick A drug-drug interactions study between reference guide should facilitate clinicians’ ability to rosuvastatin and pantoprazole after oral answer questions about the concurrent use of herbal administration in healthy male subjects medicine products and prescription medicines, thus Huguet J1,2, Gaudette F2, Bélanger F2, Dallaire enhancing patient safety. S2,4, Iglesias L2, Doucet J2, Nelson M2, Chiasson JL2,3,4, Hamet P 2,3, Turgeon J1,2 30 1Faculty of Pharmacy, University of Montreal, Role of metabolite receptor GPR91 in post- Montreal, Quebec, Canada; 2Research Centre, Centre stroke recovery 2 3 2 1 Hospitalier de l'Université de Montréal (CRCHUM), Hamel D , Sanchez M , Rivera JC , Hou X , Montreal, Quebec, Canada; 3Department of Medicine, 3 1,2,3 Bhosle V , Chemtob S University of Montreal, Montreal, Quebec, Canada; Hôpital Sainte-Justine Research Center, Department of 4Research Group on Diabetes and Metabolic 1Pediatrics and 2Pharmacology, Université de Regulation, CRCHUM, Montréal, Québec, Canada Montréal, Montréal, Québec, Canada; 3Department of Corresponding Author: [email protected] Pharmacology & Therapeutics, McGill University, Conflict of Interest: On behalf of all authors, cannot Montréal, Québec, Canada identify any potential conflict of interest. Corresponding Author: [email protected] Conflict of Interest: None declared Purpose: Rosuvastatin (ROSU) disposition involved intestinal and hepatic membrane transporters such as Background: Numerous events affecting normal OATP1B1, 1B3, 2B1, ABCG2 (BCRP) and NTCP. cerebral blood flow can cause hypoxia-ischemia (H-I), Pantoprazole (PANTO) is a potential which can lead to major neurological disabilities in infants substrate/inhibitor of ABCG2. Our objective was to and adults. There is currently a void in treatment select PANTO as a clinical probe to study the impact modalities to counter cerebrovascular injury. We have of ABCG2 inhibition on the pharmacokinetic of recently demonstrated that the Krebs cycle product ROSU. succinate, links capillary function to tissue metabolic Methods: Eight healthy White male subjects needs via the G protein-coupled receptor (GPCR) GPR91. performed a single-center, 2-period, cross-over study. The functional benefits of this coupling and its effect on They received, on 2 occasions, 7 days apart, either an post-stroke recovery are still unknown. oral dose of PANTO (40 mg) or placebo in the Objectives: We assess the role of succinate in morning followed by a single oral dose of ROSU (10 orchestrating brain angiogenesis and recovery during mg) 1 hour after. The next morning (24 hours after), post-ischemic events in the newborn. subjects were given a 2nd 40 mg dose of PANTO. Methods: To test our hypothesis the Rice-Vanucci Genotyping for 3 single nucleotide polymorphism model was employed. Succinate levels were evaluated ABCG2 C421A, SLC01B1 T521C and A388G by mass spectrometry at different time-point following (OATP1B1) was performed. Blood samples were H-I. Expression of GPR91 was determined by collected before and 0.25, 0.5, 0.45, 1, 1.5, 2, 3, 4, 6, 8, immunohistochemistry and western blot. Expression of 10, 12, 24, 26, 28, 30, 32, 48 and 72 hours after ROSU e330 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 administration. Urine was collected over 72 hours. circulation will transfer across the placenta and will be Plasma and urine samples were analyzed by toxic to both the placenta and fetus. LC/MSMS. Data were analyzed by ANOVA with Objectives: First, to determine whether formic acid repeated measures adjusting for sequence and period transfers across the placenta and is toxic to the for Cmax, AUC, CL/F and CLr. No effect boundary of placenta. Second, to determine whether folate can 80-125% for the point estimates and the 90% decrease transplacental transfer of formic acid and confidence interval (CI) for the ratio of the geometric mitigate toxicity. least-square means of all PK parameters were Methods: Dual perfusion of a single placental lobule determined. ex vivo was used to characterize the transfer of formic Results: Geometric means for Cmax, AUCinf and acid across the placenta. After a 1-hour control period, CL/F were respectively 4.3 ng/mL, 44.39 h·ng/mL and formic acid (2mM) was introduced into the maternal 3754.21 mL/min for ROSU alone, and 4.21 ng/mL, circulation with (n=4) or without folate (1uM) (n=4) 45.89 h·ng/mL and 3631.30 mL/min for and allowed to equilibrate for 3-hours. ROSU+PANTO. Ratio and 90% CI were 96.48% Results: Formic acid transferred rapidly from the (83.67% - 114.24%), 100.20% (90.41% - 111.90%) maternal to the fetal circulation and transfer was not and 99.58% (86.67% - 107.95%) for Cmax, AUCinf altered with the addition of folate. Compared to the and CL/F, respectively. Geometric means for CLr were control period, there was a significant decrease in hCG 245.82 mL/min (ROSU) and 232.24 mL/min secretion (p=0.03) after addition of formic acid. In (ROSU+PANTO). Ratio and 90% CI were 98.99% contrast, there was no significant decrease when folate (87.73% - 101.73%). However, in one subject was present in the perfusate. homozygous for variant alleles (521CC) associated Conclusions: Formic acid rapidly transfers across the with a loss of function in OATP1B1, an increase in placenta and thus has the potential to be toxic to the plasma concentrations was observed following developing fetus. Formic acid decreases hCG secretion pantoprazole administration. Cmax and AUCinf were in the placenta, which may alter steroidogenesis and respectively 8.5 ng/mL and 83.83 h·ng/mL for ROSU differentiation of the cytotrophoblasts, and this can be alone, and 10.2 ng/mL and 90.33 h·ng/mL for mitigated by folate. ROSU+PANTO. Conclusion: ROSU is an ABCG2 (BCRP) substrate 33 for its elimination through the bile. However, this study Maternal fish consumption and mercury: risk showed that block of ABCG2 (BCRP) by pantoprazole perceptions and therapeutic monitoring does not alter rosuvastatin pharmacokinetics to a Jahedmotlagh Z1,4, Schoeman K1,2, Bend JR1,2,3, significant extent in healthy volunteers with functional 2,3,4,5 activities of other membrane transporters involved in Koren G Departments of Pathology1, Physiology & the disposition of the drug. Pharmacology2 and Paediatrics3, Schulich Medicine & Dentistry, University of Western Ontario, Ivey Chair in 32 Molecular Toxicology4, London, Ontario, Hospital for Placental transfer of formic acid is rapid and Sick Children5, Toronto, Ontario decreases hCG secretion Corresponding Author: [email protected] Hutson JR1,2, Lubetsky A1, Eichhorst J3, Hackmon Conflict of Interest: None declared R1, Koren G1,2, Kapur BM1,4 1Division of Clinical Pharmacology and Toxicology, Background: While fish is rich in essential nutrients Hospital for Sick Children, Toronto, ON; 2Insitute of and women are encouraged to consume fish products, Medical Sciences, University of Toronto, Toronto, ON; fish may contain methyl mercury which is an 3Saskatchewan Disease Control Laboratory, Regina, established neurotoxin to the fetus. Consequently, there SK; 4Dept of Clinical Pathology, Sunnybrook Health are high levels of anxiety among women of Sciences Center, Toronto, Canada reproductive age regarding fish consumption. Corresponding Author: [email protected] Objectives: 1) To investigate what motivates women Conflict of Interest: None for all authors of reproductive age to avoid eating fish during their pregnancy and to understand their perceptions towards Background: Formic acid has recently been detected consuming fish; and 2) to pilot an intervention program in maternal blood and umbilical cord blood of infants in women of reproductive age to ensure mercury levels born to alcohol abusing mothers. This toxic metabolite are below the LOAEL. of methanol requires folate for detoxification. We Methods: We surveyed 100 women of reproductive hypothesize formic acid produced in the maternal age who consulted the Motherisk program about fish consumption on their perceptions regarding fish

J Popul Ther Clin Pharmacol Vol 18 (2):e315-e363; May 22, 2011 e331 © 2011 Canadian Society of Clinical Pharmacology and Therapeutics. All rights reserved. 2011 CSPT ANNUAL CONFERENCE - ABSTRACTS consumption. Subsequently we implemented a MCT1 at a greater level than MCT4. Experiments therapeutic monitoring program for women who had conducted to assess the functionality of the transporters hair mercury levels above 0.3 µg/g, the No Observable in these cell lines confirmed the lactic acid uptake in Effect Level (NOEL) for mercury for neurocognitive both of those cell lines. effects. Conclusions: Different cancer cell lines can be used as Results: The majority of women (90%) were aware of in vitro models for the transport study of lactic acid. the potentially harmful effects of fish containing high Experiments are currently underway to link MCT1 and levels of mercury. Most respondents were unable to MCT4 transport activity to drug toxicity. describe specific toxic effects. When rating the level of anxiety from 0 (none) to maximal (10), the mean rate 35 was 5, and 16 women were most worried. A pilot on 5 Respiratory depression following therapeutic women testing above the LOAEL of 0.3 mcg/g (mean administration of opioids in the operating 0.78±0.46), after diet modifications reducing fish room: an opioid pathway pharmacogenetic consumption, levels decreased significantly (to 0.33±0.22) (P<0.01). analysis Madadi P1, Sistonen J2,3, Gladdy R4, Silverman Conclusion: Women are very concerned about fish 5 5 2,3 3,6 consumption and potential fetal risk. Therapeutic G , Carvalho JC , Ross CJD , Carleton BC , 2,3 1 monitoring appears to be effective. Hayden MR , Koren G 1Clinical Pharmacology and Toxicology, Hospital for 2 34 Sick Children, Toronto, Canada; Centre for Molecular Transport of lactic acid by mct1 and mct4 in Medicine and Therapeutics, University of British Columbia, Vancouver, Canada; 3Child and Family cancer cell lines 1,2 2 1,2 Research Institute, University of British Columbia, Leung YH , Belanger F , Armstrong C , 4 5 1,2 Vancouver, Canada; Dept. of Surgery, and Dept. of Turgeon J Anesthesia and Pain Management, Mt. Sinai Hospital, 1 Faculty of Pharmacy, Université de Montréal, Toronto, Canada; 6Pharmaceutical Outcomes 2 Montreal, QC, Canada; CRCHUM, Centre de Programme, Children’s and Women’s Health Centre of Recherche du Centre Hospitalier de l’Université de British Columbia, Vancouver, Canada Montréal, QC, Canada Corresponding Author: [email protected] Corresponding Author: [email protected] Conflict of Interest: None declared Conflict of Interest: No conflict of interest to report Background: Systemic approaches are needed to Background/Objectives: A decrease in oxygen supply understand how variations in the genes associated with to mitochondria is associated with a reduction in ATP opioid pharmacokinetics and pharmacodynamics can production from pyruvate and an increased formation be used to predict clinical outcome. We present 2 cases of lactate. Accumulation of lactate, and secondly of of life threatening opioid-induced respiratory lactic acid, is associated with mild to severe depression in the operating room. Case One: The pathophysiological conditions including severe muscle patient had severe respiratory depression following 2 pain. Two monocarboxylate transporters namely, mg of subcutaneous morphine on top of intrathecal MCT1 (SLC16A1) and MCT4 (SLC16A3), are morphine administered for a Cesarean section. The involved in the transport of lactic acid in several cell patient had a history of near apnea with one dose of types. The objective of our studies was to develop cell codeine/acetaminophen (30mg/500mg respectively), models to dissect activity of these two transporters. but tolerated hydromorphone. Case Two: Life Methods: Seven different breast cancer cell lines were threatening respiratory depression occurred following cultured and harvested for RNA. RT-PCR analyses epidural morphine given at standard doses for surgical were performed to determine the relative mRNA removal of tumor. Post-operatively, the patient needed expression of the MCT1 and MCT4. For uptake only 0.6mg total of IV hydromorphone over 4 days for transport assay, cells were plated and incubated with pain management. 14 [ C] lactic acid at 37°C. The level of lactic acid Methods: Functional candidate polymorphisms in incorporated was determined by using a liquid genes involved in opioid metabolism and action scintillation analyzer. pathway (CYP2D6, UGT2B7, ABCB1, OPRM1, Results: RT-PCR results obtained allowed us to COMT) were genotyped by using SNaPshot® and identify, out of the seven breast cancer cell lines, two TaqMan® Drug Metabolism Genotyping assays or by cell lines with interesting characteristics. First, MDA- amplifying and re-sequencing the corresponding MB-231 expressed MCT4 at a much higher level than genomic regions. MCT1 while, on the other hand, SKBR3 expressed e332 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

Results: Case One: Genotype results revealed this The maximum enhancement of Gd occurred within 12 patient had an increased propensity to generate active minutes following the administration of LPA. Re- metabolites from both codeine (extensive CYP2D6 establishment to normal barrier function was apparent activity) and morphine (increased UGT2B7 activity) within 20 minutes of LPA exposure. Examination of while having a functional µ-opioid receptor system. the permeability of a large near infrared fluorescent These active metabolites are not generated with imaging agent, IRdye PEG, showed a qualitatively hydromorphone. similar accumulation profile. Case Two: Collectively, this patient appeared to have Conclusions: LPA produces a rapid and reversible increased exposure and overall sensitivity to morphine increase in brain microvessel endothelia cell and hydromorphone. Decreased ABCB1 efflux permeability. These studies indicate that administration transporter activity, in combination with low COMT of LPA in combination with therapeutic agents may be activity associated with increased sensitivity of the µ- an effective strategy to increase drug delivery to the opioid receptor system may have predisposed the brain. patient to this adverse outcome. Conclusions: An opioid pathway pharmacogenetic 37 approach along with clinical history may provide Ocular toxicity in children exposed in utero to insight into severe respiratory depressive events in antimalarial drugs: a systematic review of the patients who received therapeutic doses of opioids and literature may be useful information to mitigate future adverse Osadchy A, Ratnapalan T, Koren G events. Motherisk Program, Division of Clinical Pharmacology & Toxicology, Hospital for Sick Children, Toronto, 36 Canada Rapid and reversible enhancement of blood- Corresponding Author: [email protected] brain barrier (BBB) permeability using Conflict of Interest: The authors state no conflict of lysophosphatidic acid (LPA) interest to declare. On N, Miller DW Department of Pharmacology and Therapeutics, Background: There have been concerns regarding Faculty of Medicine, University of Manitoba, retinal toxicity in the offspring of women exposed to Winnipeg, Canada antimalarial drugs chloroquine (CQ) and Corresponding Author: [email protected] hydroxycholoquine (HCQ) during pregnancy. Conflict of Interest: None declared Objective: To systematically review the published evidence on safety of antimalarials during pregnancy Background: The delivery of drugs to the CNS is with focus on ocular toxicity in the offspring. limited due to the restrictive nature of the BBB. Methods: Ovid MEDLINE(R), EMBASE and Transient modulation of BBB permeability is one Cochrane Library databases were searched for method for enhancing drug delivery to the brain. One randomized controlled trials (RCTs) and observational potential modulator of BBB permeability is LPA. studies assessing visual function in the offspring of Objectives: Examine the use of the LPA, to transiently women exposed to antimalarials during pregnancy. increase BBB permeability Results: 12 studies with a total of 588 exposed Methods: LPA-induced alterations in brain offspring met the inclusion criteria. Of 12 studies, 2 microvessel permeability were examined in both cell were RCTs and 10 were cohort studies 5 of which were culture and whole animal models. The permeability of lacking comparison group. Methods and time of visual fluorescein labeled dextran (FDX; MW3000) was assessment varied among studies. 5 studies reported no examined using human brain microvessel endothelial clinical visual abnormalities in all cases (n= 251). In a cells, HBMEC, under control conditions and following RCT on malaria prophylaxis, visual acuity in 251 exposure to LPA (0.1-10 μM). Effects of LPA on BBB infants exposed to CQ in utero did not differ from permeability was examined in Balb/c mice using placebo group. Detailed ophthalmological examination magnetic resonance imaging (MRI) and near infrared was performed in 4 studies and normal results were fluorescence imaging techniques. reported in all children (n=59). Electrophysiological Results: Exogenous LPA produced concentration- testing using electroretinogram was performed in 3 dependent increases in FDX permeability in HBMEC. small cohorts of infants exposed to HCQ prenatally (n= Mice treated with LPA (1mg/kg) had significantly 31) and were normal in all but six infants. higher accumulation of Gadolinium contrast agent (Gd) Conclusions: The current evidence from small and compared to control mice in all regions of brain with relatively low quality studies suggests no fetal ocular the greatest increase observed in the posterior regions. toxicity of antimalarials during pregnancy. The clinical

J Popul Ther Clin Pharmacol Vol 18 (2):e315-e363; May 22, 2011 e333 © 2011 Canadian Society of Clinical Pharmacology and Therapeutics. All rights reserved. 2011 CSPT ANNUAL CONFERENCE - ABSTRACTS significance of early electroretinogram anomalies reported in other studies. Elevated midazolam levels, reported in a small subset of infants remains to be correlating with EEG recordings suggestive of established. Larger follow up studies are warranted to oversedation may be a risk factor for the development confirm low risk of ocular toxicity in children of delirium. Further data is being analyzed and will be following antenatal exposure to antimalarial available for presentation. medications. 39 38 Systematic monitoring of amiodarone therapy The Delirium Risk Evaluation and Assessment produces high efficacy and low toxicity rates of Midazolam, EEG Recording and Sleep Pollak PT, Gillis AM, Wyse DG, Burland JB, (DREAMERS) Study Arburg B Ovakim DH1,2, Bosma KJ2,3, Young GB2,4, Sen University of Calgary, Calgary, AB, Canada M2,3, Tirona R1, Kim RB1, Dresser GK1 Corresponding Author: [email protected] 1Division of Clinical Pharmacology, Department of Conflict of Interest: None declared Medicine, University of Western Ontario; 2Critical Care Western, Division of Critical Care Medicine, Background: Amiodarone, although highly effective, Department of Medicine, University of Western is renowned for potential adverse effects limiting its Ontario; 3Division of Respirology, Department of use. In clinical trials of patients receiving arbitrary Medicine, University of Western Ontario; 4Division of doses and minimal monitoring, 15-20% discontinued Clinical Neurological Sciences, University of Western amiodarone in the first year. Ontario, London, Canada Objectives: To report outcomes in patients taking Corresponding Author: [email protected] amiodarone who are managed in a specialized Atrial Conflict of Interest: None declared Fibrillation (AF) Clinic with regular monitoring, individual serum drug concentration guided Background: Critically ill patients require adequate amiodarone dose titration, and focused education. sedation to tolerate the invasive interventions necessary Methods: 60 patients, who were naïve to amiodarone for their care. Benzodiazepines are commonly used to when started in outpatient clinic, were followed for ≥ achieve sedation in the intensive care unit (ICU), 12 months. Thyroid, liver and lung function were however, they have been linked to the development of recorded at baseline, then, monitored along with annual delirium. Midazolam, a benzodiazepine commonly pulmonary metrics according to symptoms. used in ICU demonstrates significant interindividual Amiodarone dosing was adjusted using guidance from pharmacokinetic (PK) variability in these patients. serum amiodarone concentrations. Objectives: 1) To define the relationship between Results: As a mixture of persistent and intermittent AF midazolam PK and electroencephalogram (EEG) in patients, 25% were in sinus rhythm at baseline (B). critically ill patients. 2) To clarify the effect of critical After 12 mo (E) of therapy, 0% discontinued illness on midazolam pharmacokinetics. 3) To amiodarone and 90% were in sinus without limiting determine if impaired midazolam clearance is a risk adverse effects or clinically important changes in liver factor for delirium. or thyroid function (ALT B=33 vs. E=39 and FT4 Methods: Patients admitted to the ICU with sepsis and B=14.9 vs. E=18.3). Thyroid supplementation was on a continuous infusion of midazolam were screened required in 15 pts (13 already on thyroxin at baseline) for study enrolment. Upon enrolment, continuous and later 4 pts developed transient hyperthyroidism that subhairline EEG was applied and daily blood samples resolved while still on amiodarone. Some had transient were collected for plasma midazolam quantification. neurologic symptoms that resolved after loading. No Clinical and laboratory parameters were followed and pulmonary toxicity was observed. delirium onset was monitored using the Intensive Care Conclusion: Systematic monitoring and dose Delirium Screening Checklist (ICDSC). adjustment based on serum amiodarone concentrations Results: Data is currently available for five patients. provides a high rate of therapeutic success. Thyroid Patients on continuous midazolam infusions had issues were common, but no patient required maximal plasma midazolam concentrations ranging discontinuation of amiodarone for toxicity. from 176 to 472 ng/ml (mean 336 ± 118). Corresponding EEG tracings demonstrated predominance of a delta 40 wave pattern suggestive of deep sedation. Four out of 5 Hair cortisol concentrations in patients with patients had ICDSC scores suggestive of delirium. obstructive sleep apnea Comment: The midazolam concentrations observed Russell E1, Koren G1,2,3,4, Rieder MR2,5, Rotenberg suggest impaired clearance, and are higher than B6, Ferguson KA2, Uum SV2 e334 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

1Department of Physiology and Pharmacology, 41 University of Western Ontario, London, Canada; Pharmacogenetics of warfarin in children 2 Department of Medicine, Schulich School of Medicine Shaw K1,2,3, Amstutz U1,2,3, Rassekh SR4, Braun and Dentistry, University of Western Ontario, London, 4 3,5 3,5 1,2,3 3 J , Ross C , Hayden MR , Carleton BC Canada; Ivey Chair in Molecular Toxicology, 1Faculty of Medicine, Department of Paediatrics, UBC, University of Western Ontario, London, Canada; Vancouver, Canada; 2Pharmaceutical Outcomes 4Department of Clinical Pharmacology/Toxicology, 5 Programme, BC Children’s Hospital, Vancouver, Hospital for Sick Children, Toronto, Canada; CIHR- Canada; 3Child & Family Research Institute, GSK Chair in Pediatric Clinical Pharmacology, Vancouver, Canada; 4Division of Paediatric Children’s Health Research Institute, London, Canada; 6 Hematology/Oncology/BMT, BC Children’s Hospital, Department of Otolarngology – Head and Neck Vancouver, Canada; 5Department of Medical Genetics, Surgery, University of Western Ontario, London, Faculty of Medicine, Centre for Molecular Medicine Canada and Therapeutics, UBC, Vancouver, Canada Corresponding Author: [email protected] Corresponding Author: [email protected] Conflict of Interest: None declared Conflict of Interest: None declared

Background: Obstructive sleep apnea (OSA) is a Background: Warfarin is a highly effective common sleep disorder with serious cardiovascular and anticoagulant, but the large variation in dose metabolic co-morbidities. OSA patients experience a requirements between patients makes dosing repetitive collapse of the upper airway, resulting in challenging. Inadequate warfarin dosing can cause intermittent episodes of hypoxemia. The adverse serious adverse drug reactions (ADRs), such as blood effects of OSA may be mediated by increased cortisol clots or excessive bleeding. Polymorphisms in three secretion. In this model, the frequent sudden arousals genes (CYP2C9, VKORC1 and CYP4F2) have a during sleep activate the hypothalamic-pituitary- significant effect on the required warfarin dose in adrenal axis resulting in increased nighttime cortisol adults. However, the validity of these findings have not secretion, a time when cortisol secretion is normally been comprehensively assessed in children. Further very low. Hair analysis is a non-invasive tool that can paediatric studies are required to understand the provide a retrospective, integral measure of cortisol predictive factors contributing to dose variation in production over several months. children and prevent warfarin-induced ADRs. Objectives: This study will assess if hair cortisol Objectives: To determine the effect of genetic concentrations can be used as a biomarker of OSA variation in CYP2C9, VKORC1 and CYP4F2 on severity. It is hypothesized that hair cortisol content warfarin response in children, as well as the importance correlates with the severity of OSA, and successful of additional variation in genes involved in drug intervention with either CPAP or surgery will result in biotransformation and coagulation pathways that may decreased cortisol concentrations. be implicated in paediatric warfarin dosing. Methods: Patients are recruited after undergoing a Methods: We will collect a cohort of paediatric sleep study. Their hair cortisol concentrations are patients receiving warfarin therapy and test for determined with an immunosorbent assay and associations of genetic variation in VKORC1, CYP2C9 compared with their apnea-hypopnea index (AHI), a and CYP4F2 with therapeutic dose, time to stable score of the severity of a patient’s OSA. A second, international normalized ration (INR), and warfarin- post-intervention sample will be collected from induced ADRs. We will also use univariate analysis to positively diagnosed patients undergoing an test for associations between therapeutic dose and intervention. additional genes studied. Results: To date, 65 pre-intervention patients have Significance: The anticipated results of this study will been recruited, and many post-intervention hair provide insight into the genetic basis of warfarin dose collections will occur in upcoming months. A requirements in children and the potential benefits of preliminary analysis of 39 patients using a linear genetic testing in children prior to initiation of warfarin regression did not detect a correlation between cortisol therapy. A paediatric-specific genetic dosing algorithm concentration and AHI. would allow early prediction of patients at risk for Conclusion and Plan: In March, 22, post-intervention over- or underanticoagulation, and minimize the danger samples will be collected, allowing for a preliminary associated with warfarin therapy in this under-studied assessment of if cortisol concentrations decrease with population. effective interventions for OSA.

42 Paediatrics & Pharmacology, University of Montreal, Creation and assessment of adenovirus- Montreal, QC, Canada mediated drug transporter model expression Corresponding Author: [email protected] system for the prediction of pharmacokinetic Conflict of Interest: None declared profiles in humans Purpose: The importance of identifying VEGF- Sugiyama D, Teft WA, Knauer M, DeGorter MK, independent pathways in pathological angiogenesis is Tirona RG, Kim RB increasingly recognized as a result of emerging drug Division of Clinical Pharmacology, Department of resistance to anti-VEGF therapies. Human T- Medicine, The University of Western Ontario, London, Lymphocyte-Derived Microparticles (LMPs) Canada significantly inhibit angiogenesis in several ocular Corresponding Author: [email protected] neovascularization (NV). Both pigment epithelium- Conflict of Interest: None declared derived factor (PEDF) and the neurotrophins (NT) low- affinity p75NTR receptor have shown antiangiogenic Introduction: Predicting the in vivo role and effects of effects. Our study is designed to determine how LMPs functional genetic polymorphisms to observed PK profile modulate the pro and antiangiogenic of drugs in humans is important for optimal drug therapy. microenvironments in choroidal angiogenesis. Currently, IVIVE (in vitro to in vivo extraporation) Methods: Antiangiogenic effects of LMPs were algorithms are widely utilized for the prediction of drugs determined by using rat model of choroidal explants. which are metabolized by CYP enzymes, but not LMPs were produced by treatment of human T- transporters. Therefore, our goal is to create an in vitro lymphocytes with actinomycin D. Cell viability (MTT transporter expression system capable of expressing assay), proliferation ([3H]-thymidine DNA multiple drug transporters simultaneously so that the cell- incorporation), migration assays and apoptosis, were based system better reflects human organs such as the tested in cell lines. Choroidal expression of VEGF, liver. Accordingly, we have cloned a number of hepatic PEDF, nerve growth factor (NGF) and p75NTR were bile acid and drug uptake transporters into an adenovirus- demonstrated by Western blots and RT- PCR. based expression construct and tested the efficiency of Results: Choroidal NV was suppressed by more than transporter expression in a number of cell lines. 50% after 72h of LMPs treatments. LMPs targeting Methods: Adenovirus constructs containing uptake acted on multiple cell types important for choroidal transporters such as NTCP and members of the human angiogenesis, such as vascular endothelial cells (inhibit OATPs were constructed and transport activity, as well HREC cell proliferation by 55%), and RPE cells as, cell viability assessed in HeLa, MDCKII, LLC-PK1 (ARPE19). At a molecular level, LMPs regulated and Caco-2 cell lines. neurotrophins and their receptors expression both in Results: In terms of cell viability, LLC-PK1, Caco-2 vitro and in vivo. Inhibition of p75NTR abolished the and Hela were able to tolerate MOI (Mutiplicity of antiangiogenic effect of LMPs. Infection) of 1000, but lower for MDCKII at MOI 300. Conclusions: LMPs are important candidate for Transport function at the same MOI was the greatest in antiangiogenic therapy. PEDF and neurotrophin HeLa cells. Interestingly, MDCKII cells exhibited induction by LMPs may be of therapeutic value in higher transport activity compared to LLC-PK1 cells treating ocular neovascular diseases. Our data when expressing NTCP, but LLC-PK1 was better when demonstrate that choroidal tissues have the capacity to expressing OATP2B1. synthesize neurotrophins, and that various stimulations Conclusions: Our findings suggest a number of cell can up-regulate gene and protein expression of lines can be transduced to express hepatic transporters neurotrophins. using the adenoviral expression system. Therefore, our in vitro system has the potential serve as a 44 physiologically relevant model for predicting in vivo PK. Dehydroepiandrosterone alters retinol levels and expression of retinol related proteins 43 Takitani K, Miyazaki H, Tamai H Choroidal antiangiogenic effects of lymphocyte- Department of Pediatrics, Osaka Medical College, Japan Corresponding Author: [email protected] derived microparticles are mediated through Conflict of Interest: None declared PEDF and neurotrophin receptor p75NTR signalling pathways Background: Dehydroandrosterone (DHEA) and its Tahiri H, Yang C, Gagnon C, Picard E, Chemtob S, sulfate DHEA-sulfate ester (DHEA-S) are the most Hardy P abundant adrenal steroids in humans. However, the e336 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 physiologic roles of DHEA and DHEAS have not been responses to ethanol and caffeine compared with clearly defined. High levels of DHEA have been heterozygous mice. reported to be associated with decreased risk of Methods: To examine ethanol sensitivity, we tested cardiovascular disease and there has been speculation loss of righting response (LORR) duration after about their possible role in the aging process. In animal injection (i.p.) of ethanol (3.6g/kg; 20% v/v in saline). experiments, DHEA and DHEA-S have beneficial To examine caffeine sensitivity, alterations in effects to obesity, diabetes, oncogenesis, locomotor activity were monitored after injection of atherosclerosis, and memory. Vitamin A is essential for caffeine (25 mg/kg, i.p.). vision, embryonic development reproduction, Results: In behavioural assays, transgenic mice immunity, and growth. The affect of DHEA to retinol showed a greater response to ethanol and a reduced status has not been reported previously. response to caffeine than wild type littermates, but no Objectives: In this study, we examined the retinol significant differences between heterozygous and status in rats administered DHEA and investigated the homozygous transgenic mice were detected. expression of retinol related proteins including lecithin Conclusion: These data indicate that the increase in retinol acyltransferase (LRAT) and beta-carotene ENT1 function between wild type and heterozygous 15,15´ monooxygenase (BCM) genes, which are mice is greater than the increase in ENT1 function metabolic enzymes of retinol and beta-carotene between heterozygous and homozygous transgenic respectively. mice. Therefore, homozygous mice do not offer a Methods: Wistar rats (four weeks, male) were significant advantage over heterozygous mice for assigned to two groups: a control group and a DHEA studies of ENT1 regulation of adenosine levels and group fed the standard rat chow containing 0.4 % adenosine dependent behaviours. (wt/wt) DHEA, and fed for two weeks. Results: Retinol levels of both plasma and liver in 46 DHEA administered rats are decreased compared with Cardiac specific over-expression of membrane- controls. Hepatic BCM and LAT gene expression is associated human stem cell factor promotes significantly decreased in DHEA administered rats. epicardial activation post myocardial Expression of both enzymes may affect circulatory infarction retinol status. Conclusions: DHEA and DHEA-S are 1 3 1 3 1,2,3 widespread as supplements for anti-aging. However, Xiang F-L , Lu X , Liu Y , Liu M , Feng Q Departments of 1Physiology and Pharmacology, and we should be aware that excess of DHEA intake might 2 3 affect fat-soluble retinol status. Medicine, University of Western Ontario, Lawson Health Research Institute, London, Canada 45 Corresponding Author: [email protected] Conflict of Interest: None declared Behavioural effects of enhanced expression of equilibrative nucleoside transporter 1 in mice Background: Myocardial infarction (MI) is one of the Sun C, Kost S, Xiong W, Parkinson F leading causes of death worldwide. We recently Department of Pharmacology & Therapeutics, Faculty demonstrated that the cardiac specific over-expression of Medicine, University of Manitoba, Winnipeg, of membrane-associated human stem cell factor (MA- Canada hSCF) improves cardiac function and survival post-MI. Corresponding Author: [email protected] Epicardium derived cells (EPDCs), a population of Conflict of Interest: None declared cardiac stem cells which are essential for embryonic heart development, has recently been shown to be able Background: Adenosine is a neuromodulator that to be activated and involved in cardiac repair post-MI. permeates cell membranes via nucleoside transporters. Objective: The aim of the present study was to Mice with neuronal expression of human equilibrative investigate the effects of cardiomyocyte-specific over- transporter 1 (hENT1) have been generated (Parkinson expression of MA-hSCF on epicardial activation post- et al., 2009 J. Neurochem. 109:562-572). Expression of MI. We hypothesized that cardiac specific over- hENT1 was associated with increased ataxic effects of expression of MA–hSCF promotes epicardial ethanol and reduced stimulatory effects of caffeine, activation in mice post MI. two drugs that act at least in part through adenosine Methods/ Results: Wild-type (WT) and the inducible signalling mechanisms. cardiac-specific MA-hSCF transgenic (hSCF/ tTA) Objectives: The present study examined the mice were subjected to MI. Activated EPDCs were hypothesis that mice homozygous for the hENT1 increased in hSCF/tTA epicardium compared to WT transgene have significantly different behavioural mice (P<0.05) 3 days post MI as determined by Wt1 staining. E13.5 WT EPDCs were cultured and infected

J Popul Ther Clin Pharmacol Vol 18 (2):e315-e363; May 22, 2011 e337 © 2011 Canadian Society of Clinical Pharmacology and Therapeutics. All rights reserved. 2011 CSPT ANNUAL CONFERENCE - ABSTRACTS with Ad-hSCF or Ad-EGFP. Proliferation was Conclusions: Our finding gives evidence that clearly significantly enhanced in Ad-hSCF infected EPDCs as CAL induced relaxation in rabbit Aorta rings via determined by cell counting (P<0.05). Moreover, a involvement in the mobilization of Ca2+ from ER as trans-well system was employed to evaluate the well as from Ca+2 channels. Our results might led to migration of E13.5 EGFP+ EPDCs. Neonatal WT new therapeutic application of CAL in cardiovascular cardiomyocytes were cultured and infected with Ad- diseases. hSCF or Ad-LacZ in the lower compartment of the + trans-well. EGFP EPDCs were seeded in the upper 48 compartment. Twenty-four hours later, EGFP signal in Synthesis and anticonvulsant properties of 1- the bottom well was significantly increased in the Ad- (amino-N-arylmethanethio)-3-(1-substituted hSCF infected group compared to Ad-LacZ (P<0.05). benzyl-2, 3-dioxoindolin-5-yl) urea derivatives Conclusions: Cardiomyocyte-specific over-expression Alam MS1, Siddiqui N1, Stables JP2 of MA-hSCF promotes the activation of EPDCs post- 1 MI. Over-expression of MA-hSCF enhances the Department of Pharmaceutical Chemistry, Faculty of proliferation and migration of EPDCs. Pharmacy, Hamdard University, Hamdard Nagar, New Delhi-110062, India, 2Epilepsy Branch, National Institute of Neurological Disorders and Stroke, NIH, Rockville, MD 20892, USA Corresponding Author: [email protected] POSTERPRESENTATIONS Conflict of Interest: None declared THURSDAY- MAY 26, 2011 Introduction: There is continuing demand for new 47 anticonvulsant agents as several of the currently Calcium involved in the vasorelaxant effect of available antiepileptic drugs (AEDs) have been convolvulus arvensis L extract on rabbit aorta associated with severe side effects and fail to control rings seizures in about 30% of epileptic patients. Study Al-Aghawani W, Abu Jayyab A revealed that isatin is a privileged lead molecule for Faculty of Pharmacy, International University for designing potential bioactive agents. Science & Technology, Daara, Syria Objectives: Considering extensive applications of Corresponding Author: [email protected] isatin moiety in medicinal chemistry, an attempt has Conflict of Interest: None declared been made to synthesize amino-N-arylmethanethio urea derivatives containing isatin moiety that are Background: Our previous finding suggests that comparatively more efficacious and safer than the calcium plays an important role in the relaxant effect of currently used anticonvulsant agents. Convolvulus arvensis L (CAL) extract in rabbit's aorta. Methods: A series of new 1-(amino-N- Objective: As a result, further investigation is needed arylmethanethio)-3-(1-substituted benzyl-2, 3- to determine calcium role in the CAL induce dioxoindolin-5-yl) urea (5a-p) were prepared. The relaxation. pharmacological testing is done by National Institute of Methods: CAL extract was added cumulatively to the Neurological Disorders and Stroke (NINDS), USA rabbit aorta pre-contracted with high K+ Krebs under Anticonvulsant screening program (ASP). The solution. Also another procedure was performed using Phase I pharmacological screening comprised MES, Ca2+-free Krebs where aorta ring exposed to scPTZ and neurotoxicity. Some of the selected Phenylephrine (PE) as a control group. When the compounds were tested for their activity at 6 Hz model. Results: The most potent anticonvulsant phasic contractions reached a plateau, CaCl2 was added to the bath, causing a tonic contraction. This procedure compounds found to be were 5f, 5h, 5i, 5k and 5l. In effectively explain whether the relaxant action of CAL MES test, compound 5h and 5i showed marked was due to intracellular Ca release from the protection at 100 and 300 mg/kg dose. In sc.PTZ endoplasmic reticulum (ER)or extracellular Ca2+- screening compounds 5h and 5i were active at dose of release from cell surface calcium channels. The 300 mg//kg. In 6 Hz screening procedure was repeated in the same tissue after compounds 5h and 5i showed significant protection pretreatment of extract or Diltiazem. and emerged as lead compounds. Results: CAL significantly decreased vessels Conclusions: Compounds 5h and 5i emerged as a lead contraction induced by high concentration of K+. Also, compound compared to the standard drugs when they the CAL extract inhibited both calcium release from were subjected to preliminary anticonvulsant the ER and the entry of calcium through Ca2+ channels. screenings. They showed marked lower neurotoxicity

e338 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 and therefore a higher protective index. These can be 50 regarded as strong candidates for future investigations. Endothelial microparticles promote oxidative stress and inflammation in cultured mouse 49 aortic endothelial cells: role of epidermal Bone marrow cells migrate to the retina and growth factor receptor effect angiogenesis in a mouse model of oxygen Burger D, Thompson CS, Touyz RM induced retinopathy Kidney Research Centre, Ottawa Hospital Research Blais M1, Rivera JC1, Madaan A1, Sapieha M3, Institute, University of Ottawa, Canada Chemtob S1,2 Corresponding Author: [email protected] 1Research Center, Sainte-Justine Hospital, Montreal, Conflict of Interest: None declared Quebec, Canada; 2Department of Ophthalmology and Pharmacology, University of Montreal, Montreal, Background: Microparticles (MPs), submicron Quebec, Canada; 3Department of Ophthalmology, fragments of cellular membranes shed from Maisonneuve-Rosemont Hospital Research Centre, stressed/damaged cells are found in the plasma of University of Montreal, Montreal, Quebec, Canada healthy individuals with levels increased in vascular Corresponding Author: [email protected] injury. Elevated plasma MP levels correlate with Conflict of Interest: None declared vascular dysfunction and predict future cardiovascular events. However, whether MPs themselves contribute Background: The retina is a metabolically active to endothelial dysfunction is unclear. tissue requiring a large amount of oxygen. Under Objectives: We tested the hypothesis that endothelial hypoxic conditions, neovascularization occurs and can MPs, through EGFR, influence endothelial cell (EC) have severe negative repercussions on the retina. function by increasing EC oxidative stress and Retinopathy of prematurity (ROP), the leading cause of stimulating pro-inflammatory responses. infant blindness, exemplifies the detrimental effects Methods: Endothelial MPs were isolated from the neovascularization can cause. These premature media of cultured mouse aortic ECs by centrifugation newborns acquire vaso-obliteration of micro vessels in and quantified by flow cytometry. ECs were treated 5 the retina, followed by a pathological with endothelial MPs (10 /ml) and effects on ROS neovascularization once the retinal metabolic demand generation (DHE HPLC), pro-inflammatory signaling increases. Oxygen induced retinopathy (OIR) is an (adhesion molecule expression) and inflammatory animal model that replicates both phases of ROP and is responses (macrophage adhesion) were examined. achieved by exposing newborn mouse pups to 80% Results: Endothelial MPs significantly increased oxygen from post-natal day 7 to 12. There have been production of superoxide anion in ECs (~2-fold, many advances in stem cell research regarding P<0.05) after 4 hours and expression of the cellular coronary and renal revascularization demonstrating a adhesion molecules PECAM (~3-fold, P<0.05), and possible role for stem cells in angiogenesis. VCAM (~3-fold, P<0.05) after 8 hours. Treatment with 5 Objective: Utilizing bone marrow derived stem cells, endothelial MPs (10 /ml) significantly increased we aimed to repopulate the retina with normal vessels macrophage adhesion to ECs after 8 hours (P<0.05). which are affected in the OIR model. Examination by confocal microscopy suggested a Methods: Two different cell types were isolated from surface interaction between ECs and MPs. mouse bone marrow: lineage negative (Lin-) and Additionally, western blot analysis identified the mesenchymal stem cells (MSC). These cells were then epidermal growth factor receptor (EGFR) ligand HB- injected into the vitreous of OIR mice. EGF. We therefore tested the hypothesis that MPs Results: Mouse retinas collected at P17 from both promote oxidative stress and inflammation through MSC and Lin- injected mice demonstrated cell EGFR activation. Co-treatment with the EGFR migration to the inner retina. Furthermore, MSC inhibitor gefitinib (1 µM), blocked MP-induced injected mice retinas showed reduced oxidative stress and inflammation. neovascularization and vaso-obliteration where as Lin- Conclusions: In summary, we demonstrate that injected retinas did not result in significant endothelial MPs are pro-oxidative and pro- revascularization. inflammatory in ECs. These effects appear to be Conclusion: MSCs migrate to the retina in mice mediated through surface interaction and stimulation of having undergone the OIR model and promote proper EGFR. Thus MPs may be more than just biomarkers of vascular repair. These results suggest that MSCs play a vascular injury and may themselves contribute to role in angiogenesis and could have a therapeutic use in endothelial dysfunction. retinopathy of prematurity.

51 1Centre de Recherche, Institut de Cardiologie de 2 Hair analysis as a tool for estimating child Montréal, Montréal, QC, Canada, Pharmacologie, 3 exposure to environmentally relevant Médecine, Université de Montréal, Montréal, QC, polybrominated diphenyl ethers (PBDEs) Canada. Carnevale A1,2, Aleksa K1,3, Koren G1,2,4 Corresponding Author: [email protected] Conflict of Interest: None declared 1Division of Clinical Pharmacology and Toxicology, The Hospital for Sick Children, Toronto, Canada; Background: Calcium signaling is fundamental for 2Department of Pharmacology and Toxicology, endothelial functions and an alteration in the regulatory University of Toronto, Toronto, Canada; 3School of mechanism can lead to cardiovascular pathologies. Pharmacy, University of Waterloo, Waterloo, Canada; Recently, spontaneous calcium oscillations localized 4Ivey Chair in Molecular Toxicology, University of within the myoendothelial junctions have been Western Ontario, London, Canada identified and named calcium pulsars. Corresponding Author: Calcium/Calmodulin Kinase II (CaMKII) is a potential [email protected] target for Ca2+ pulsars involved in endothelial Conflict of Interest: None declared function. Indeed, CaMKII has recently been suggested to play an important role in endothelial function and Background: Polybrominated diphenyl ethers dysfunction. CaMKII is characterized by its unique (PBDEs) are chemicals that are added to a variety of ability to interpret intracellular calcium oscillations consumer products as flame retardants. They are frequency into specific outcomes. Therefore, alteration persistent in the environment and have been detected in in endothelial CaMKII functions might be related with wildlife and in humans. The human body burden in intracellular calcium oscillations such as calcium North America is among the highest in the world. pulsars. The aim of this study is to investigate the Some studies have reported higher levels of PBDEs in relationship between CaMKII and calcium pulsars and children, than in their mothers, as measured in serum. it’s alteration in hypertension. Acute and chronic PBDEs are structurally similar to other persistent stimulation of calcium pulsars with phenylephrine organic pollutants, some of which can potentially appears to increase CaMKII activation in endothelium interfere with endocrine pathways. This is of concern from murine mesenteric arteries. Moreover, CaMKII since childhood is an ongoing period of growth and translocates following calcium pulsars stimulation. development. Although found in clusters in hypertensive mouse, Objectives: To establish a method to quantify PBDEs CaMKII is homogeneously distributed in the in the hair of children (newborn-age 16) as a biomarker endothelium from CTRL mice. qPCR experiments of long-term systemic exposure. revealed that all four CaMKII isoforms expression (α, Methods: A method has been developed using gas β, γ and δ) are significantly diminished (25-57%) in chromatography with mass spectrometry (GC/MS). mesenteric arteries from hypertensive mice. In Fifty mg of children’s hair is incubated overnight at summary, the increase of calcium pulsars frequency in 40°C with 4N HCl and hexane (4:1) to extract PBDEs. hypertensive mice correlates with CaMKII’s Samples are eluted from 2g NaSO : 2g Florosil SPE 4 distribution in endothelial cells, suggesting their columns with 8mL of hexane. Samples are then activation by calcium pulsars. Therefore, an altered analyzed by GC/MS for PBDE congeners; BDE-28, - relationship between CaMKII and calcium pulsars can 47, -99, -100, -153, -154, -183 and -209. potentially be involved in endothelial dysfunction Results: The total amount of PBDEs varied among associated to hypertension. samples. Several congeners could be detected in newborn hair. BDE-209 was present in some samples. The greatest variability was seen with congeners BDE- 53 47 and BDE-99. Development and pharmacological Conclusions: This method offers a noninvasive characterization of a new class of peptidic technique for assessing chronic exposure to PBDEs in urotensin II antagonists children, and to examine correlations between systemic Chatenet D1,2, Nguyen QT3, Létourneau M1,2, exposure and adverse developmental outcomes. Dupuis J3,4, Fournier A1,2 1Laboratoire d’études moléculaires et 52 pharmacologiques des peptides, Université du Québec, Impact of hypertension on endothelial CaMKII INRS – Institut Armand-Frappier, Ville de Laval, Québec, Canada; 2Laboratoire International Associé isoforms from mesenteric arteries 3 Charbel C1,2, Toussaint F1, Blanchette A1, Samuel de Champlain (INSERM – INRS); Montreal Villeneuve LR1, Ledoux J1,3 Heart Institute, Université de Montréal, Montréal, e340 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

Québec, Canada; 4Department of Medicine, Université against ethical, religious, and psychosocial issues of de Montréal, Montréal, Québec, Canada pregnancy termination. Corresponding Author: [email protected] Objective: To identify the medical, ethical, cultural Conflict of Interest: None declared and spiritual aspects involved in the process of decision-making regarding the termination of a wanted Background: Known antagonists of the urotensin II pregnancy after exposure to a teratogen and to provide (UII) receptor (UT) are of limited utility for guidelines for optimal management. investigating the pathophysiological role of UII due to Methods: A case discussion took place including poor potency and limited selectivity. Clinical Pharmacology fellows and staff, graduate and Objectives: Design, synthesis and pharmacological post graduate students, counselors, obstetricians, evaluation of new UII antagonists. bioethics faculty, and representatives of all religious Methods: The pharmacological properties of a novel Chaplains of the Hospital for Sick Children, in order to UT antagonist, [Bip4]URP, was investigated in vitro discuss and find better ways to support women in their and in vivo using several bioassays. decision making process. Results: Competitive binding assays demonstrated the Summary: 1) Woman should be explained her legal ability of [Bip4]URP to fully displace the radioligands rights in decision-making, including the fact that 125I-hUII and 125I-URP from human recombinant UT. abortion is legal and free in Canada. 2) One-on-one In ex vivo studies, various concentrations of counseling on fetal risk following exposure should be [Bip4]URP did not produce any significant rightward provided based on evidence-based information. 3) The shift of the hUII concentration-response curve, but the role of the father and family members in supporting the maximal response to hUII was not attainable. woman’s decision making should be stressed. 4) Interestingly, a slight but non significant rightward Religious scholars should be involved for spiritual shift, along with a non significant reduction of efficacy, support. 5) In the event of termination, long term was observed with URP. Supporting these support from medical, religious, psychosocial, and observations, dissociation experiments revealed the family members should be provided. propensity of this compound to accelerate the Conclusion: Collaboration of healthcare providers, dissociation rate of hUII but not URP, suggesting the bioethics and chaplaincy representatives, and family presence of two binding pockets in close vicinity. In members, is essential to reduce the women’s moral vivo, [Bip4]URP had no effect on the biphasic response distress when faced with a decision of pregnancy induced by URP but significantly reduced the termination after teratogen exposure. hypotensive activity, while keeping intact the pressor effect induced by hUII. These results demonstrated its 55 selectivity against hUII agonistic effect. Benzodiazepine use in the Grand-Duchy of Conclusion: The results demonstrated the ability of 4 Luxembourg from 1995 to 2006: proposed [Bip ]URP to reduce the efficacy of hUII- but not definitions of high-dosage use and abuse URP-induced vasoconstriction. Moreover, in vivo Cloos J-M1, Bocquet V2, Rolland-Portal I3, Koch studies support the in vitro pharmacological profile 3 4 described above, making [Bip4]URP as the first P , Chouinard G 1ZithaKlinik, Luxembourg; 2CRP-Santé, Luxembourg; peptidic analog of a new class of urotensinergic 3 antagonists. Contrôle Médical, Ministry of Social Security, Luxembourg; 4Department of Psychiatry and Medicine, 54 McGill University Health Center, Montréal, Canada Corresponding Author: [email protected] Ethics involved in termination of a wanted Conflict of Interest: None declared pregnancy in reproductive toxicology Chaudhry S, Koren G, Nulman I Background: Prevalence rates of benzodiazepine The Motherisk Program, Division of Clinical intake vary among studies, mainly due to definitions of Pharmacology and Toxicology, The Hospital for Sick benzodiazepine use, abuse, lengths of observation Children, Toronto, Canada period and samples of subject. Corresponding Author: [email protected] Objective: To examine short- and long-term use of Conflict of Interest: None declared benzodiazepines in a large national sample. Methods: A 12-year population-based study was Background: A serious conflict is created when a conducted in the country of Luxembourg by looking at woman must decide whether or not to terminate a benzodiazepine prescriptions for all insured subjects in wanted pregnancy after exposure to a teratogen. She the national health system from 1995 (n =387,862) to must weigh the fetal/child risks following exposure 2006 (n =449,972). Patterns of benzodiazepine use and

J Popul Ther Clin Pharmacol Vol 18 (2):e315-e363; May 22, 2011 e341 © 2011 Canadian Society of Clinical Pharmacology and Therapeutics. All rights reserved. 2011 CSPT ANNUAL CONFERENCE - ABSTRACTS characteristics of benzodiazepine users were studied. Thymidine incorporation and MTT assay were used as Results/Conclusion: The overall number of Defined an OPC proliferation and cell viability indexes, Daily Dose (DDD) per 1000 insured subjects per day respectively. OPC protein extracts were resolved by was 82.9; the five most prescribed benzodiazepines Western Blotting. were lormetazepam, lorazepam, alprazolam, Results: We found that Phen alone increased OLP bromazepam and loprazolam. Alprazolam was the only proliferation and potentiated the effects of IGF-1. In benzodiazepine showing a threefold increase in its addition, IGF-1-stimulated signaling pathways were annual prescribed volume during the study period. further increased by Phen treatment including Akt, S6 Subjects having had at least one benzodiazepine ribosomal protein and ERK. prescription (n=236,263) in the 12-year study period, Conclusion: Overall, our first results suggest a were divided into 3 groups: 1) a “short term delivery” potential role of IGF-1 on OLG proliferation and group (34.9%) with benzodiazepine prescription ≤ 3 survival under PTEN inhibition. months; 2) a “discontinuous delivery” group (38.4%); and 3) a “continuous delivery” group (26.7%) of 57 WITHDRAWN subjects who never stopped taking benzodiazepines Recurrence and long-term outcome of Stevens- once prescribed. High-dose use was defined using a Johnson Syndrome and toxic epidermal new formula to calculate potential abuse, and a necrolysis in children constant number of high dose users were found 1,2,5,6,8 1,5 8 throughout the study period: 5.3% of all users (0.9% of Finkelstein Y , Soon GS , Acuna P , George M8, Pope E1,4,5, Ito S1,3,5,6, Shear NH3,6,7, Koren all subjects) had a yearly benzodiazepine intake higher 1,3,5,6 8 1,3,5 than the maximum recommended dosage. G , Shannon MW , Garcia-Bournissen F 1Department of Pediatrics and Divisions of 2Emergency Medicine, 3Clinical Pharmacology and Toxicology and 56 4 IGF-1 effects on oligodendrocyte survival and Section of Dermatology, Hospital for Sick Children; Departments of 5Pediatrics, 6Pharmacology and proliferation are potentiated through PTEN Toxicology, and 7Dermatology, University of Toronto, inhibition Toronto, ON, Canada; 8Clinical Pharmacology Unit, De Paula M, Almazan G Division of Emergency Medicine, Children’s Hospital Department of Pharmacology & Therapeutics, McGill Boston, Harvard Medical School, Boston, MA, USA University, Montreal, Canada Corresponding Author: [email protected] Corresponding Author: Conflicting Interests: None declared [email protected] Conflict of Interest: None declared 58 Potential use of synthetic kinin B1 receptor Background: Myelin is a multilayer specialized membrane that wraps nerve fibers and insulates them peptide agonists as permeability enhancers for facilitating the communication between neurons. improving drug delivery to malignant brain Myelin destruction causes neurological impairments tumours 1 2 1 1 such as those observed in patients with multiple Gobeil F Jr , Côté J , Savard M , Bovenzi V , sclerosis. Oligodendrocytes (OLGs) are cells Dubuc C1, Lepage M2, Fortin D3 responsible for producing myelin in the central nervous 1Dept. of pharmacology; 2Dept. of nuclear medicine system. Insulin-like growth factor-1 (IGF-1) is and radiobiology; 3Dept. of surgery, Univ. de essential for OLG growth, maturation and survival. Sherbrooke, Canada IGF-1 binds to its receptor to activate the Corresponding Author: PI3K/Akt/mTOR cascade which mediates protein [email protected] synthesis, important for OLG progenitors (OLPs) Conflict of Interest: None declared growth. PTEN (phosphatase and tensin homologue deleted on chromosome 10) is the major negative Background: Some evidence suggests that kinin B1 regulator of PI3K/Akt/mTOR pathway. receptors (B1R), an inducible G protein-coupled Objectives: Our aim is to assess whether an inhibitor receptor (GPCR), can regulate blood-brain barrier of PTEN, potassium bisperoxo oxovanadate (Phen), (BBB) permeability, including that of brain tumours. could be used as pharmacological tool to increase or Objectives: 1) To analyze expression of B1R in brain potentiate IGF-1 effects on OLP survival and tumour of syngeneic F98 glioma-implanted Fischer proliferation. rats. 2) To determine whether the 8 9 Methods: Primary cultures of OPC were prepared SarLys[DPhe ]desArg -bradykinin (NG29), a stabilized from the brains of newborn Sprague-Dawley rats. kinin B1R agonist, modulates blood brain barrier e342 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

(BBB) function, thereby improving delivery of Objectives: To determine whether acyclovir is a macromolecules directly into brain tumours. substrate for the human BCRP. Methods: Expression of B1R (mRNA, protein) in Methods: Cellular accumulation studies were brain normal and tumour tissues was determined by conducted to determine whether acyclovir is a substrate RT-PCR, Western blot, and IHC. Effects of NG29 for the human BCRP. Transfected human embryonic were monitored by non invasive MRI with kidney (HEK293) cells [containing the full-length Gadolinium-based contrast agents Gd-DTPA (0.5 kDa) human ABCG2 gene encoding the wildtype ABCG2 and Gadomer (17 kDa) (T1-weighted imaging), and by amino acid sequence] were exposed to [8-14C] IHC of endogenous albumin (~66 kDa). acyclovir in the presence or absence of the BCRP Results: Preferential expression of B1R at tumour cells inhibitor, fumitremorgin C (FTC) for 2 hours. and surrounding tumour vasculature were detected. Intracellular [8-14C] acyclovir accumulation was then Intracarotid infusion of NG29, in contrast to natural assessed using a liquid scintillation counter. LysdesArg9-bradykinin, elevated brain distribution and Results: The results illustrated that acyclovir is a uptake profiles of intravenous contrast agents within rat substrate for human BCRP. In the presence of FTC, glioma and brain surrounding. These effects were dose- there was a 5-fold increase (p<0.05) in the intracellular dependent, reversible (lasting< 2h), and were blocked accumulation of acyclovir. by B1R antagonist R892 and non-selective COX Conclusions: The study is the first to illustrate that inhibitor indomethacin, but not by B2R antagonist acyclovir is a substrate for the human BCRP. The HOE140 and NO-synthase inhibitor L-NA. Consistent results suggest that BCRP may play a significant role with MRI data, immunostaining for extravasated in the renal clearance of acyclovir, and contributes to albumin at the invasive tumor edge was increased in the further understanding of the tubular transport of NG29-treated rats. acyclovir. Future studies are required to determine the Conclusion: Our results document a novel GPCR affinity of the transporter for the antiviral agent. signalling mechanism for promoting transvascular delivery into brain tumour, involving possibly COX 60 byproducts. They also underline the potential value of The effect of N-acetylcysteine on the synthetic B1R agonists as selective tumour BBB antitumour efficacy of ifosfamide in a mouse permeabilizers for local delivery of different-sized xenograft model therapeutics at (peri)tumoral sites. Hanly L1, Chen N1, Rieder M1,3,4, Yeger H6, Koren G1,2,3,5,7 59 1Department of Physiology and Pharmacology, Acyclovir is a substrate for the human breast University of Western Ontario, London, Ontario, cancer resistance protein (BCRP) 2 1,2 1,3 1,2 Canada; Ivey Chair in Molecular Toxicology, Gunness P , Aleksa K , Koren G Schulich School of Medicine and Dentistry, University 1 Division of Clinical Pharmacology and Toxicology, of Western Ontario, London, Ontario, Canada; The Hospital for Sick Children, Toronto, Ontario, 3Department of Paediatrics, University of Western 2 Canada; Graduate Department of Pharmaceutical Ontario, London, Ontario, Canada; 4CIHR-GSK Chair Sciences, Leslie Dan Faculty of Pharmacy, University in Paediatric Clinical Pharmacology, Children’s Health 3 of Toronto, Canada; School of Pharmacy, University Research Institute, London, Ontario, Canada; 5Division of Waterloo, Ontario, Canada of Clinical Pharmacology and Toxicology, Hospital for Corresponding Author: [email protected] Sick Children, Toronto, Ontario, Canada; 6Divisions of Conflict of Interest: None declared Hematology/Oncology, Hospital for Sick Children, Toronto, Ontario, Canada; 7Department of Background: The renal transport mechanisms of Pharmacology, Faculty of Medicine, University of acyclovir have not been fully elucidated. The human Toronto, Toronto, Ontario, Canada breast cancer resistance protein (BCRP) is a transporter Corresponding Author: [email protected] that is widely expressed in human tissues, including the Conflict of Interest: None declared kidney. Studies illustrate that, in mice, BCRP mediates the transport of acyclovir into breast milk. It is Background: It is estimated that 82% of children will plausible that acyclovir is a substrate for human BCRP, survive childhood cancer, up from 50% decades ago. and hence, the transporter may be actively involved in While promising, this increased life span has brought the tubular efflux of acyclovir. The role of human awareness of the high potential that these patients may BCRP in the transport of acyclovir has not been develop late effects, conditions secondary to cancer previously evaluated. therapy. Nephrotoxicity is one such late effect, affecting children treated with the chemotherapy drug

J Popul Ther Clin Pharmacol Vol 18 (2):e315-e363; May 22, 2011 e343 © 2011 Canadian Society of Clinical Pharmacology and Therapeutics. All rights reserved. 2011 CSPT ANNUAL CONFERENCE - ABSTRACTS ifosfamide, commonly used to treat pediatric solid Background: The currently recommended guidelines tumours. While effective, in children it is associated to minimize the incidence of NTD are well established; with a 30% risk of developing nephrotoxicity, with 5% serum/plasma folate >7 ng/mL (15.9 nmol/L) or RBC of this group developing Fanconi’s syndrome. N- folate concentrations >906nmol/L. However, there is acetylcysteine, as synthetic thiol that is currently used controversy regarding the relationship between serum clinically in children, has successfully mitigated this folate/folic acid supplement use and pregnancy renal toxicity in both cell and rodent models. However, outcome. before this treatment can be realized clinically, we Objective: To evaluate the relationship between serum must demonstrate that it does not interfere with the folate/folic acid supplement use and pregnancy antitumour efficacy of ifosfamide. Our objective is to outcome. compare the efficacy of ifosfamide with and without Methods: As part of the Prospective Cohort Study of concurrent n-acetylcysteine therapy in an the MOCEH, 1220 pregnant women were enrolled. immunocompromised mouse xenograft model. Female Information about folic acid supplementation, maternal Swiss nu/nu mice will be injected with Ewing’s serum folate levels in the 1st & 3rd trimester of Sarcoma tumour cells. They will receive 60mg/kg pregnancy, and pregnancy outcomes, were obtained. injections of ifosfamide (3 days), with or without Comparison of pregnancy outcomes between women 1.2g/kg injections of n-acetylcysteine (concurrently who received prenatal folic acid supplementation and and for + 3 days). Tumour volumes will be measured. those who did not receive supplementation was Preliminary data assessing the efficacy of ifosfamide conducted. A post-hoc analysis was performed during concurrent n-acetylcysteine therapy in vitro, in comparing pregnancy outcome a) between women with relevant tumour cell lines (rhabdomyosarcoma and blood folate levels <7 ng/mL vs. >7 ng/mL, and b) neuroblastoma) indicate n-acetylcysteine has no effect. between women with <20 ng/mL vs. >20 ng/mL folate We anticipate similar findings with our rodent model levels. (to be completed before May). Upon anticipated Results: The mean and median level of serum folatel results, these findings will demonstrate the clinical were 11.7 ng/mL and 9.6 (95% CI 3.3-26.9), applicability of n-acetylcysteine for ifosfamide-induced respectively. Overall 32.9% (n= 401/1220) of early nephrotoxicity and strengthen support for a clinical pregnant women were at <7ng/mL folate. Only 29.8% trial. (n= 363/1220) of pregnant women took multivitamins with folic acid during the 1st trimester of pregnancy. 61 Serum folate level was higher in the FA supplement The relationship between folic acid group than in the no-FA supplement group. No supplementation and serum folate level in early differences in pregnancy outcomes were observed a) between women who received folic acid pregnancy and pregnancy outcomes: MOCEH supplementation and those who did not, and b) between (Mothers and Children’s Environmental women with blood folate levels <7 ng/mL vs. >7 Health) study ng/mL. Although most preterm deliveries <34 weeks 1,2 3 4 3 5 Hong S-C , Ha E-H , Han J-Y , Park H , Ha M , were observed in the group of serum folate <20 ng/mL, Kim Y6, Hong Y-C7, Nava-Ocampo AA1, Koren there was no statistical significance. Other pregnancy G1 outcomes such as preeclampsia, gestational diabetes, 1Motherisk Program, Hospital for Sick Children, placenta previa and premature rupture of membrane Toronto, ON, Canada, 2Korea University Medical remained similar between groups. Center, Seoul, South Korea , 3Department of Preventive Conclusions: Although most preterm deliveries was Medicine, School of Medicine, Ewha Medical observed in the low serum folate levels (<20 ng/mL), Research Center, Ewha Womans University, 4Korean there was no significant relation between serum folate Motherisk Program, Cheil General Hospital and level/folic acid supplementation and pregnancy Women's Healthcare Center, Seoul, South Korea, outcomes. This is possibly due to unsufficient sample 5Department of Preventive Medicine, Dankook size, and should be repeated with larger cohorts. University College of Medicine, 6Department of Occupational and Environmental Medicine, Ulsan 62 University Hospital, University of Ulsan College of NK1 receptor antagonists modulate the effects 7 Medicine, Department of Preventive Medicine, Seoul of immunosuppressive drugs on T cells National University College of Medicine activation: an in vitro study Corresponding Author: [email protected] Jizi K1, Welman M1, Maghni K1 Conflict of Interest: None declared

1Centre de recherche de l’hôpital du Sacré-Coeur de metric to use in determining the optimum dose for this Montréal, Université de Montréal, Montréal, QC, group of patients. We conducted a review of available Canada literature to determine availability of pharmacokinetic Corresponding Author: [email protected] data in obese children. Conflict of Interest: None declared Methods: A review of available adult and paediatric obesity pharmacokinetic literature was conducted. Introduction: Cyclosporin A (CsA) and Tacrolimus Results: Definitions of obesity are even more (FK506) both inhibit T cell activation to prevent or challenging in children than in adults. Many measures reduce graft reject. Rapamycin also regulate T cells of body composition are available, but have not been function by repressing cell proliferation. Aside from extensively verified. Whereas absorption seems not to their therapeutic efficiency, these molecules have be affected, distribution, metabolism and elimination several side effects. An option to reduce their adverse are known to be in various extend. Plasma protein effects is to decrease the dose of immunosuppressive levels and binding are comparable to non-obese, and drugs by combining it with another molecule. The NK1 effects on regional blood flow are unclear. Evidence receptor (NK1R) expressed on T cells may be a good suggests that hydrophilic drugs whose Vd in normal- candidate because this receptor under activation by its weight subjects is small should be administered ligand, the substance P (SP), regulates T cells function. according to ideal body weight, and not total body The goal of this study is to demonstrate the potential weight. CYP450 enzyme activity is altered for the 2E1 efficacy of combining a NK1R antagonist (L-733,060) isoform, but for others results are conflicting. with CsA, FK506 or rapamycine. Glomerular filtration rates are mostly similar, but this Methods: Jurkat T cells were incubated with is unclear for tubular reabsorption. rapamycin (1; 0,5 and 0,1 nM) in presence of a NK1R Conclusions: There is a lack of available antagonist L-733,060 (1; 5 and 10 uM). The level of pharmacokinetic data for the determination of optimal phosphorylation of the S6 ribosomal protein (S6R) was dosing schedules for obese children, and adults alike. measured by flow cytometry. Jurkat cells were also incubated with CsA (1, 5 and 10 ng/ml) or FK506 64 (0,01; 0,05 and 0,1 ng/ml) with the L-733,060. The Pharmacogenetics of fatal paediatric modulation of IL-2 production by T cells was obstructive sleep apnea cases in response to measured by ELISA. codeine: additional cases Results: The combination of rapamycin with L- 1 2 3 4 733,060 did not affect the level of phosphorylation of Kelly L , Rieder M , Veenstra F , Madadi P , Ross C5, Sistonen J6, Koren G7,8 the S6R protein. On the other hand, the combination of 1 CsA or FK506 with L-733,060 significantly reduces Schulich School of Medicine and Dentistry, University of Western Ontario, London, Canada; IL-2 production in stimulated T cells. 2 Conclusion: The NK1R signalling pathway is Department of Pediatrics and Medicine, Children's Hospital of Western Ontario, University of Western implicated in the production of IL-2 in activated T cells. 3 Combining a NK1R antagonist with immunosuppressive Ontario, London, Canada; Grey Bruce Health Services Hospital; 4Department of Pharmacology, Hospital for drugs may be an interesting alternative to reduce 5 therapeutic doses without affecting therapy efficacy. Sick Children, University of Toronto; Department of Medical Genetics, Centre for Molecular Medicine and Therapeutics, University of British Columbia; 6Centre 63 for Molecular Medicine and Therapeutics, University Pharmacokinetics of obesity in childhood – a of British Columbia; 7Motherisk Program, Hospital for review Sick Children, Toronto; 8Ivey Chair in Molecular Jong GW, Koren G Toxicology, University of Western Ontario, London, Division of Clinical Pharmacology & Toxicology, Canada Hospital for Sick Children, Toronto Ontario, Canada Corresponding Author: [email protected] Corresponding Author: [email protected] Conflict of Interest: None declared Conflict of Interest: None declared Background: The primary treatment for pediatric Introduction: The obesity epidemic is widely obstructive sleep apnea (OSA) is removal of the tonsils recognized as one of the biggest health threats of the and adenoids (adenotonsillectomy). Codeine is modern age. It is not uncommon for clinicians in prescribed for the pain associated with this procedure. primary or secondary care to treat illness in an obese Codeine is a prodrug which requires conversion to child, which will require treatment with a drug for morphine via the highly polymorphic cytochrome P450 which there is no guide regarding which body size (CYP) 2D6 enzyme to elicit its analgesic properties. A

J Popul Ther Clin Pharmacol Vol 18 (2):e315-e363; May 22, 2011 e345 © 2011 Canadian Society of Clinical Pharmacology and Therapeutics. All rights reserved. 2011 CSPT ANNUAL CONFERENCE - ABSTRACTS functional gene duplication causes increased morphine Methods: A systematic review and meta-analysis were production in the ultra-rapid metaboliser (UM) conducted. MEDLINE, Embase and SCOPUS were phenotype. In 2009, we reported a case of fatal searched from inception to May 2010 with the respiratory depression in a 2yr old male post- following search terms “folic acid”, “C677T”, adenotonsillectomy with toxic morphine levels. Post- methylenetetrahydrofolate reductase”, “colorectal mortem analysis revealed bronchopneumonia and a cancer”, “colonic neoplasms”, “rectal neoplasms”. CYP 2D6 UM phenotype. We report now 2 additional Observational studies in adult populations were Ontario cases. In September 2010 we recorded a near included that reported MTHFR C677T variants, fatal case of respiratory depression in a 3yr old female defined levels of folic acid and the risk of colorectal OSA patient. She arrived in hospital comatose with cancer. toxic blood morphine levels despite using Results: Out of 633 records, 28 studies met our recommended codeine doses. Upon mechanical inclusion criteria. Preliminary results indicate that the ventilation and standard naloxone treatment she fully summary risk estimate for case control studies recovered. Salivary genetic analysis revealed that she comparing MTHFR CC to CT was 0.92 (CI 95% 0.83- was a CYP 2D6 UM. A third, fatal case was 1.01) with some heterogeneity. The summary risk encountered in November 2010, again, with toxic estimate for case control studies comparing MTHFR blood morphine levels after taking only recommended CC to TT was 0.82 (CI 95% 0.71-0.95) with some doses. This case of respiratory depression occurred in a heterogeneity. 4 yr old male. This patient had a previous history of Conclusion: This meta-analysis supports the asthma and was identified as a CYP 2D6 UM. We association between the TT genotype and a reduced hypothesize that the combination of an UM CYP 2D6 risk of colorectal cancer. Further analysis is required to phenotype and an existing respiratory condition in provide insight into whether this risk is further toddlers with OSA increases the risk for central modified through folic acid intake levels. nervous system depression upon standard codeine administration. 66 Transcriptional regulation and function 65 organic anion transporting polypeptide 2B1 MTHFR C677T polymorphism and the risk of splice variants colorectal cancer: a systematic review and Knauer MJ1, Girdwood AJ1, Kim RB1,2, Tirona meta-analysis RG1,2 1,2 1,3 1 Kennedy DA , Stern SJ , Sarkar M , Adams- 1Department of Physiology and Pharmacology; Webber T1, Koren G1,2,3 2Department of Medicine, Division of Clinical 1The Motherisk Program, The Hospital for Sick Pharmacology, Lawson Health Research Institute, The Children, Toronto, ON, Canada; 2Leslie Dan Faculty of University of Western Ontario, London, Ontario, Pharmacy, University of Toronto, Toronto, ON, Canada Canada; 3Department of Pharmacology and Corresponding Author: Toxicology, University of Toronto, Medical Sciences [email protected] Building, Toronto, ON, Canada Conflict of Interest: None Corresponding Author: [email protected] Conflict of Interest: None declared Background: The human Organic Anion Transporting Polypeptide 2B1 (OATP2B1) is a membrane Background: Folates are water soluble B vitamins that transporter that facilitates the cellular uptake of a are cofactors in thymidine and purine synthesis and number of endogenous compounds and drugs. DNA methylation pathways. OATP2B1 is expressed in several tissues including the Methylenetetrahydrofolate reductase (MTHFR) is key small intestine, liver, kidney and skeletal muscle. in the remethylation of homocysteine to methionine. In Recently, it has been shown that differential promoter 1995 a variant of the MTHFR enzyme, genotype TT, usage in tissues results in the expression of several was identified with reduced activity versus CC, causing OATP2B1 splice variants which utilize 5 distinct first an accumulation of homocysteine and higher rates of exons but share common subsequent exons. These thymidine synthesis. Individuals with this variant are splice variations are expected to encode either a full thought to be at a reduced risk for colorectal cancer. length or truncated protein missing 22 amino acids Objective: This meta-analysis examines whether a from the N-terminus. Since little is known about relationship exists between the variants of MTHFR OATP2B1 splice variants we investigated the relative C677T gene, folic acid intake and the risk of colorectal expression of the splice variants in key tissues cancer. responsible for drug absorption and elimination, as e346 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 well as the transport function of the truncated variant. context of renal dysfunction and concomitant use of the Methods/Results: Using variant-specific polymerase macrolide antibiotic, clarithromycin, which is known to chain reaction, both the predicted full length and inhibit P-glycoprotein-mediated efflux mechanisms of truncated forms of OATP2B1 were detected in liver, digoxin. Both drugs were discontinued. The patient kidney and small intestine, albeit in differing was hospitalized and was discharged ten days later. proportions. With heterologous expression in cultured Health care providers need to be vigilant of this cells, we compared the transport kinetics (Vmax and potential drug-drug interaction. A focused literature Km) of the two forms of OATP2B1. Using cell based review will be presented. reporter assays we determined that HNF4α was able to transactivate transcription of the truncated variant but 68 not the full length form. Importantly, we demonstrate Drug use reviews (DUR) in non institutional that the truncated variant was capable of transporting settings the known OATP2B1 substrates, estrone sulfate and Levine MAH, Syed U rosuvastatin. Centre for Evaluation of Medicines, St. Joseph’s Conclusion: These findings indicate that differential Healthcare and Dept. of Clinical Epidemiology & regulation of OATP2B1 splice variant expression in Biostatistics, McMaster University, Hamilton, Canada tissues could contribute to variation in drug response. Corresponding Author: [email protected] Conflict of Interest: None declared 67 Digoxin toxicity precipitated by clarithromycin Background/Objective: Drug use review (DUR) has use: case presentation and review of the become an increasingly popular component in budget literature activities conducted by drug benefit programs that Lee CYW1, Marcotte F1, Crépeau J1, Giraldeau operate in the non institutional environment. There is G1, Tardif J-C1, Koren G2, Juneau M1 great diversity in the methods used to conduct drug use 1Montreal Heart Institute, Université de Montréal, reviews, and each method has different strengths and Montreal, Quebec, 2University of Toronto, Toronto, weaknesses. This review was designed to determine the Ontario circumstances under which these methods are used. Corresponding Author: [email protected] Methods: A literature search was conducted in Conflict of Interest: None declared MEDLINE, EMBASE and CINAHL databases to identify original studies that assessed prescription drug Background: An 83-year-old man with a past history use in a community, published between January 2007 of hypertension, dyslipidemia, myocardial infarction, and February 2010. Each article was assessed for (a) and left ventricular dysfunction presented with a recent data source i.e., patient, pharmacy or prescriber; (b) history of fatigue, dyspnea on exertion, cough, nausea, drug issue of interest; (c) method used for data accrual. loss of appetite and lightheadedness. Three days prior Results: 587 articles involving 632 DUR strategies to his presentation, the patient had been started on fulfilled the criteria for inclusion in the analysis. 81% clarithromycin 500 mg p.o. b.i.d. for treatment of were retrospective studies. The source of the possible pneumonia. His cardiac medications included prescription drug use information was obtained from digoxin 0.125 mg p.o. q.d., spironolactone 25 mg p.o. pharmacy or dispensing claims data 52%, the patient q.d., captopril 25 mg p.o. b.i.d., pravastatin 40 mg p.o. 27% and physician records 21%. The most common q.d., isosorbide mononitrate 30 mg p.o. q.d., goal was the assessment of drug use patterns by furosemide 60 mg p.o. q.d., bisoprolol 2.5 mg p.o. q.d., population demographics (n=151). Measuring and potassium 20 mmol p.o. q.d. Due to severe nausea, adherence and misuse of prescription by patients was the patient presented to the Emergency Department second (n=136) and physician adherence to following completion of his fifth dose of prescription guidelines or recommendations was third clarithromycin. On examination, he appeared unwell. (n=120). When data was collected at the patient level Vital signs were stable. Cardiorespiratory examination the elapsed time between drug use and the survey was did not show signs of decompensated heart failure. more than one month 33% of the time, and in 40% of Blood tests demonstrated a white cell count of 13.6 x studies the time period was not specified; drug use 109/L, potassium 5.2 mmol/L, creatinine 184 µmol/L information was reported by a proxy 11% of the time. (creatinine was normal 3 weeks before), and digoxin Conclusion: Most DUR studies were conducted levels of 4.6 nmol/L (5 hours post-dose) and 4.7 retrospectively using prescription dispensing claims nmol/L (18 hours post-dose). Ventricular extrasystoles data to address simple quantitative questions. Studies were observed during monitoring. The presentation of designed to address questions pertaining to the quality this patient was consistent with digoxin toxicity in the of prescribing often used patient reported data to obtain

J Popul Ther Clin Pharmacol Vol 18 (2):e315-e363; May 22, 2011 e347 © 2011 Canadian Society of Clinical Pharmacology and Therapeutics. All rights reserved. 2011 CSPT ANNUAL CONFERENCE - ABSTRACTS information that would not be available from greater benefit from alternative anti-cancer treatments, administrative databases but these were conducted vincristine-induced neurotoxicity could potentially be using weaker methodologies. avoided. Additionally, the modified neurotoxicity grading scale and accurate symptom classifications 69 serves to facilitate improvements in the monitoring of Investigating the genetic causes associated with vincristine-induced neurotoxicity. incristine-induced neurotoxicity in children Loo TT1-3, Ross CJD4,5, Rassekh SR6, Hayden 70 MR4,5, Carleton BC1-3 Development of an in vitro system for the 1 functional study of oatp transports of statins University of British Columbia Faculty of Medicine, 1,2 1,2 2 2 Department of Paediatrics; 2Pharmaceutical Outcomes Lu J , Armstrong C , Belanger F , Gaudette F , 1,2 Programme, BC Children’s Hospital; 3Child & Family Turgeon J Research Institute, Vancouver; 4UBC Centre for 1Faculty of Pharmacy, Université de Montréal, Molecular Medicine and Therapeutics; 5UBC Faculty Montreal, QC, Canada; 2CRCHUM, Centre de of Medicine, Department of Medical Genetics; Recherche du Centre Hospitalier de l’Université de 6Paediatric Hematology/Oncology/BMT, BC Montréal, Montreal, QC, Canada. Children’s Hospital Corresponding Author: Corresponding Author: [email protected] [email protected] Conflict of Interest: None declared Conflict of Interest: No conflict of interest to report

Background: Vincristine is a highly effective Background/Objectives: OATP1B1 and OATP1A2 oncology drug that is considered to be the standard are known transporters of statins such as rosuvastatin. treatment against many types of cancers. However, the A mutation in these membrane transporters or drug- clinical utility of vincristine is significantly limited by drug interactions could modify plasma and intracellular debilitating adverse drug reactions (ADRs), such as concentrations of statins which can lead to cellular neurotoxicity, and more specifically, peripheral toxicity. The goal of this study was to evaluate the neuropathy. uptake of rosuvastatin by its transporters. Objectives: To identify novel predictive genomic Methods: OATP1B1 was cloned into the pIRES-EGFP markers and/or clinical factors that will determine an vector. Site-directed mutagenesis was used to generate individual’s susceptibility to vincristine-induced the loss-of-function V174A mutant, which was neurotoxicity. confirmed by sequencing. Stable cell lines were created Methods: DNA samples and detailed clinical data in HEK293 cells, and selection was done by G418 were collected through the Canadian treatment, followed by selection by Fluorescence Pharmacogenomics Network for Drug Safety Activated Cell Sorting (FACS). OATP1B1 and (CPNDS), an active ADR surveillance network OATP1B1-V174A expressing cells were then plated on encompassing 13 Canadian paediatric hospitals. ADRs Poly-Lysine treated 6-well plates, and then incubated were characterized according to a modified version of in the presence of increasing concentrations of the Common Terminology Criteria for Adverse Events rosuvastatin for various time points. OATP1A2 was (v4.03), and stratified by severity, location, and type of cloned into the pRetroX-Tight-Pur retroviral vector. neuropathy. Patient samples were genotyped for single HeLa cells were transiently transfected by this plasmid nucleotide polymorphisms (SNPs) associated with drug or the empty vector as the control and the uptake assay absorption, distribution, metabolism, and excretion. A was performed similarly as above. separate panel containing functional and tagging SNPs Results: The uptake of rosuvastatin was observed in of genes that are specifically involved in vincristine’s the presence of the wild type OATP1B1 in as little as 5 mechanism of action was also developed. minutes. However, the uptake of rosuvastatin in Results: Within CPNDS, 26.3% of paediatric cancer OATP1B1-V174A was not observed, confirming that patients on vincristine therapy suffered from this mutant is non-functional. Also, the uptake of vincristine-induced neurotoxicity (305 cases and 854 rosuvastatin was greater in OATP1A2 transfected cells controls). The neurotoxicity cases have been than those transfected with the empty vector. Controls preliminarily divided into categories of peripheral, were performed to ensure that the background signal of autonomic, and central neuropathy. rosuvastatin was minimal. Conclusions: This study aims to establish the causality Conclusions: Stable cell lines as well as transiently of vincristine-induced neurotoxicity in order to assess transfected cells expressing OATP transporters can be the risk-benefit ratio of utilizing vincristine for each used as in vitro models for functional studies. patient. By identifying individuals who would derive a e348 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

71 1Centre de recherche en rhumatologie et immunologie, Stereoisomers of naringenin as pleiotropic, Centre Hospitalier Universitaire de Québec and selective inhibitors of cytochrome P450 Department of Microbiology-Infectious Disease and Immunology, Université Laval, Québec QC, Canada isoforms 2 Lu WJ1, Flockhart DA1, Caccamese S2 Department of Biochemistry, University of Colorado Health Sciences Center, Denver, Colorado 80262, 1Department of Pharmacology and Toxicology, Indiana U.S.A. Institute for Personalized Medicine, Indiana University Corresponding Author: School of Medicine, USA, 2Dipartimento di Scienze [email protected] Chimiche, Università di Catania, Italy Conflict of Interest: The authors declare no conflict of Corresponding Author: [email protected] interest. Conflict of Interest: None declared Background: Maximakinin, a natural 19-residue Background: Naringenin is the predominant flavonone peptide from the skin of the amphibian Bombina in grapefruit, and has been suggested to be a bioactive maxima (DLPKINRKGPRPPGFSPFR), incorporates antioxidant, free radical scavenger and potential the full sequence of bradykinin (BK) at its C-terminus adjunct to treatment of Hepatitis C in humans. with a hydrophilic N-terminal extension. As a putative Interactions between naringenin and the cytochrome venom component, it may stimulate BK B receptors P450 system have been of interest since the first 2 (B Rs) in a distinct manner relative to the fragile demonstration that grapefruit juice reduced cytochrome 2 mammalian agonist BK. P450 3A activity. The effects of naringenin on other Methods: Radioligand binding assays ([3H]BK to B R- cytochrome P450 isoforms have been less carefully 2 green fluorescent protein (GFP) conjugate, investigated. In addition, the stereoisomers of [3H]enalaprilat to angiotensin converting enzyme naringenin have not been separated and purified before, (ACE)) were applied in HEK 293(a) cells to compare and so the stereoselectivity of naringenin’s effects has the affinity of maximakinin to that of BK at not been characterized. We isolated pure naringenin characteristic molecular targets. Human umbilical vein enantiomers and used them to test the ability of S-, R-, contractility and imaging/downregulation/signalling and racemic naringenin to inhibit a series of key drug studies of B R-GFP in HEK 293 cells were other metabolizing cytochrome P450 isoforms in vitro. We 2 applied assays of the effects of the peptides at natural determined the IC values for each naringenin 50 or recombinant B Rs. preparation using in vitro incubations with recombinant 2 Results: Maximakinin is an agonist of the BK B R human cytochrome P450 isoforms. We also tested the 2 with a 7-20 fold lesser potency, but a prolonged (≥ 12 ability and stereoselectivity of naringenin to inhibit h) duration of action relative to BK (ERK MAP kinase cytochrome P450-mediated drug metabolism in human activation, c-Fos induction in HEK 293 cells). liver microsomes. Naringenin was able to inhibit Maximakinin displaced [3H]enalaprilat binding from CYP2C9, CYP2C19 and CYP19 with IC values 50 recombinant ACE much less effectively than BK. below 5µM. No substantial inhibition of metabolism by Unlike BK, maximakinin induced the internalization of CYP2B6 or CYP2D6 was observed at concentrations the fusion protein B R-GFP and the downregulation of up to 10µM. The S-enantiomer exhibited higher 2 this construction over a 12-h stimulation period, fully inhibitory potency than the R-enantiomer for reproducing the effect of synthetic inactivation- CYP2C19 and CYP19, while the R-enantiomer was resistant B R agonists. more potent as an inhibitor of CYP2C9. Chiral 2 Conclusions: Maximakinin has little affinity for ACE, flavonones like naringenin are difficult to separate into a major ectopeptidase that inactivates BK, and is their enantiomeric forms, but stereoselective effects further resistant to other proteases present in the may be observed that impact clinical activity. endosome. It is a natural kinin sequence that elicits a Inhibition of specific drug metabolizing enzymes by prolonged signalling, a possible basis for a venomous naringenin observed in vitro may be exploited to action in nature and for drug development in the understand pharmacokinetic changes seen in vivo. laboratory. 72 73 Objective pharmacological properties that may Ethyl glucuronide as a biomarker of alcohol turn a vasoactive hormone into an animal consumption during pregnancy toxin: differences between bradykinin and 1,2 1,3 1,2 Matlow J , Aleksa K , Koren G maximakinin 1 1 1 2 Division of Clinical Pharmacology and Toxicology, Marceau F , Bawolak M-T , Gera L The Hospital for Sick Children, Toronto, Ontario,

Canada; 2Graduate Department of Pharmaceutical 74 Sciences, Leslie Dan Faculty of Pharmacy, University Cocaethylene as a biomarker of alcohol and 3 of Toronto, Toronto, Ontario, Canada; School of cocaine co-consumption in human hair Pharmacy, University of Waterloo, Waterloo, Ontario, Natekar A1,2, Aleksa K1,3, Koren G1,2,4,5 Canada 1Division of Clinical Pharmacology & Toxicology, Corresponding Author: [email protected] Hospital for Sick Children, Toronto, ON; 2Department Conflict of Interest: None declared of Pharmacology & Toxicology, University of Toronto, Toronto, ON; 3School of Pharmacy, Background: Alcohol consumption during pregnancy University of Waterloo, Waterloo, ON; 4Department can lead to Fetal Alcohol Spectrum Disorder (FASD), of Physiology & Pharmacology, University of Western which manifests itself in the form of physiological Ontario, London, ON; 5Ivey Chair in Molecular defects, mental retardation and neurobehavioural Toxicology, University of Western Ontario, London, deficits, the latter of which is rarely diagnosed. Since ON maternal self-reports are often unreliable, a biomarker Corresponding Author: [email protected] of alcohol use during pregnancy is needed to accurately Conflict of Interest: None declared determine fetal exposure and risk for FASD. While Fatty Acid Ethyl Esters (FAEEs) are current Background: Cocaethlyene (CE) is a metabolite of biomarkers of exposure, the introduction of a second cocaine formed only during cocaine and alcohol co- biomarker that can be tested alongside FAEEs consumption. It is pharmacologically active, promises to greatly increase the sensitivity and prolonging cocaine-related effects. specificity of analytical screens. Ethyl glucuronide Objective: To determine if CE can be used as a (EtG) is a direct metabolite of ethanol that has been biomarker in hair testing to indicate alcohol and detected in the meconium, or first stool of life, of cocaine co-consumption. infants born to mothers who consumed alcohol during Methods: We used liquid-liquid extraction and solid- pregnancy. Whether this detected EtG was formed by phase extraction to isolate cocaine and its metabolites the maternal liver and crossed the placenta, by the fetal from hair, as well as fatty acid ethyl esters, a direct liver after ethanol crossed the placenta, or by the biomarker of alcohol consumption. The compounds placenta itself remains unknown. concentrations were analyzed and determined using Objectives: To determine if, and to what extent EtG GC-MS. crosses the human placenta, and to measure placental Results: Out of 516 individuals who tested positive formation and degradation of EtG. for cocaine, 336 individuals were confirmed cocaine Methods: Placentae from consenting women users. Of these, CE was detected in 73 individuals. undergoing elective Caesarian section at St. Michael’s From those, only 23 had alcohol testing requested, and Hospital in Toronto, Ontario will be taken to the on- 15 tested positive for chronic alcohol abuse. The site perfusion laboratory. One g/mL EtG will be fraction of cocaine that was converted to CE ranged added to the maternal reservoir and maternal and fetal from 84% at low cocaine concentrations, to about 4% samples will be taken over a 3 hour period to determine at the higher cocaine concentrations. In a few cases, if and to what degree EtG is crossing the human there were difficulties in identifying CE when placenta. EtG will be quantified using HS-SPME GC- compared to benzoylecgonine, another cocaine MS. Placental metabolism of ethanol to EtG and EtG to metabolite, as the two metabolites have similar ethanol will be investigated using placental retention times on GC-MS. microsomes. Kinetic parameters (Vmax, Km) will also Conclusions: At this stage of our research, about 22% be determined to help elucidate the role of the placenta of confirmed cocaine users were positive for CE, in formation and degradation of EtG. indicating alcohol co-consumption. However, other Results: To date the perfusions are being carried out studies have found that, on average, about 62% of and the level of EtG in the perfusate will be determined confirmed cocaine users also consume alcohol. It is using HS-SPME GC-MS with a limit of sensitivity of 1 possible that hair accumulation is dose dependent and ng/vial. does not identify low alcohol use. Conclusions: We have developed a HS-SPME method for detecting EtG in maternal and fetal perfusate 75 samples. This will allow us to accurately quantify the transfer of EtG across the human placenta. Child neurodevelopment following in-utero exposure to maternal azathioprine: preliminary results Nulman I, Barrera M The Hospital for Sick Children, University of Toronto e350 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

Corresponding Author: [email protected] Background: Methotrexate is a very effective Conflict of Interest: None declared treatment for both the induction and maintenance of remission in Crohn’s disease. It is administered Background/Objective: Azathioprine (AZA) is an subcutaneously due to concerns about drug absorption immunosuppressant drug commonly used by pregnant in this patient population. Furthermore, significant women. Its effects on the fetal central nervous system inter-individual variability in its oral bioavailability has remain unknown. The objective was to assess child been demonstrated in patients with rheumatoid neurocognitive development following in-utero arthritis. exposure to AZA for maternal Inflammatory Bowel Objectives: To compare the pharmacokinetics of oral Disease (IBD), and to compare the results to those of and subcutaneous methotrexate in patients with three comparison groups. Crohn’s disease. Methods: Mother/child pairs: Group 1-exposed to Methods: A total of three patients with stable Crohn’s AZA, Group 2-exposed to corticosteroids; Group 3- disease have been enrolled thus far. Each patient exposed to other IBD medications, Group 4-exposed to received an upper endoscopy to assess for evidence of non-teratogens. Matching criteria: gender and age at upper gastrointestinal Crohn’s disease, followed by two testing (children); age (mothers). Using standardized pharmacokinetic (pk) studies – oral (PO) methotrexate psychological tests, mother/child pairs were assessed and subcutaneous (SC) methotrexate. During each on Full Scale, Verbal and Performance IQs. study day, the patients received either PO or SC Results: No significant differences were found among methotrexate followed by plasma collection at 11 pre- the 4 groups of children (3-12 years) in Full Scale IQ specified time points over a 24 hour period. (Group 1=109.2±8.1, Group 2=108.4±12.0, Group Methotrexate plasma drug concentrations were then 3=109.9±12.9, Group 4=113.9±13.7), Verbal and obtained using a sensitive mass spectrophotometer. Performance IQs. Women on corticosteroids Results: Three patients have been enrolled in this experienced more bleeding episodes during gestation ongoing study. All three had normal upper endoscopies (50%vs.3.7%vs.22.9% in AZA group and other with normal duodenal biopsies. The mean half-life was medications group, respectively). Children exposed to 2.8 hours and 3.2 hours for PO and SC methotrexate, corticosteroids had significantly shorter gestational respectively. The mean time to maximum plasma ages, lower birth weights and spent more days in concentration was 1.2 hours and 0.5 hours PO and SC NICU. The only significant predictor of these methotrexate, respectively. The area under the curve outcomes was corticosteroid group affiliation. Later ratio (PO/SC) was 0.821. There were no adverse child health parameters (growth, number of infections events. and other health problems) did not differ among the Conclusions: This unique study demonstrates that the groups pharmacokinetic parameters for PO and SC Conclusions: There were no associations found methotrexate were comparable in three patients with between AZA exposure, maternal disease Crohn’s disease. Oral methotrexate would be a more complications during pregnancy, and children’s convenient, safer, and less expensive option for these cognitive performance, which is reassuring. However, patients. no statistical differences in IQ may be a power issue. The adverse outcomes related to corticosteroids can be 77 explained by maternal disease activity. The study Cardiac effects of hyperthyroidism prevented remains in progress. by concomitant amiodarone administration during mistaken thyroid supplementation 76 Pollak PT Pharmacokinetic profiles for oral and University of Calgary, Calgary, AB, Canada subcutaneous methotrexate in patients with Corresponding Author: [email protected] Crohn’s disease Conflict of Interest: None declared Patel V1,2, Chande N1, Ponich T1, Schwarz UI2, Dresser GK2, Tirona RG2, Kim RB2, Gregor JC1 Background: Hypo-hyper-thyroid states occur more 1Division of Gastroenterology, Department of often in pts taking amiodarone. Its bi-iodinated Medicine; 2Division of Clinical Pharmacology, structure increases iodine stores in pts and inhibits Department of Medicine; University of Western conversion of T4 to T3. It takes >1 year for this iodine Ontario, London, Ontario to leave the body after stopping drug. Corresponding Author: [email protected] Objectives: To illustrate that not stopping amiodarone Conflict of Interest: None declared when thyroid abnormalities are diagnosed can have beneficial effects.

Methods: A 53-year-old man treated for atrial whether a patient obtains serum antibody against fibillation (AF) was monitored regularly (EKG, influenza virus after treating with peramivir. thyroid, liver, and serum drug monitoring). At 30 mo Materials /Methods: A multicenter and open-label of therapy, he remained asymptomatic in sinus rhythm, clinical trial without a control group was conducted in but with elevated freeT4 (42 pmol/L - ULN 25 children with influenza virus infection during the 2009 pmol/L). He was prescribed methimazole 5 mg TID pandemic A (H1N1) influenza epidemic in Japan to and continued amiodarone. evaluate the efficacy and safety of peramivir. Peramivir Results: By mistake, liothyronine 5 µg TID was was given intravenously at a dosage of 10 mg/kg (600 dispensed without detection for 57 d. Despite this, the mg maximum) once daily. Nine patients (4 m/o - 15.5 pt remained asymptomatic with HR <70 for 53 d before y/o, median: 1.2 y/o, male/female: 3/6) were enrolled having breakthrough AF when freeT4 reached 93 (3.7 in our facility and were treated with peramivir within x ULN). Total T3 showed a comparatively minor 48 hours (5.6 - 28.6 hours, median: 16.1 hours) after increase to 3.1 (1.1 x ULN of 2.8 nmol/L). developing fever. Eight patients had single dose and Conclusion: Although this pt went without prescribed one (4 m/o) had second dose due to persistent fever. antithyroid treatment, he suffered no hyperthyroid Blood samples were provided before and after the symptoms for 53 d. It appears that amiodarone administration, and serum was separated from each mitigated the effects of rising T4 by preventing a sample and analyzed for the titre of antibody against parallel rise in T3 and maintaining sinus rhythm <70. the 2009 pandemic A (H1N1) influenza virus strain in After 3 mo of methimazole, freeT4 normalized and the hemagglutination inhibition (HI) test. remains at 20. Hyperthryoidism with amiodarone needs Results: Ten samples from seven patients were careful monitoring and treatment, but is usually limited obtained before and next day of the administration to 2-8 mo duration. Stopping amiodarone does not lead (within 60 hours after the onset), and all of them to rapid depletion of iodine stores. However, showed negative HI titres less than 1: 10. Four samples continuing amiodarone at effective doses may prevent from four patients were provided 5 - 7 days after the many of the cardiovascular consequences of onset; two showed HI titres 1: 10 and the other two hyperthyroidism. showed less than 1: 10. Nine samples from each patient 1 - 2 months after onset were investigated; all showed 78 positive HI titres (1: 40 – 1: 640, median: 1: 160). Serum antibody titres against influenza virus Conclusions: Although peramivir produces rapid in pediatric patients after treating with recovery from influenza virus infection, it does not peramivir seem to interfere with obtaining antibodies in blood. Tanaka T1,2, Nagai Y1 1Department of Pediatrics, Hitachi, Ltd., Hitachinaka 79 General Hospital; 2Department of Pediatrics, Shizuoka Personalizing tamoxifen therapy for breast Kosei Hospital cancer patients 1 2 2 2 Corresponding Author: Teft WA , Dingle B , Younus J , Vandenberg T , [email protected] Brackstone M2,3, Kim RB1,2,4 Conflict of Interest: None declared 1Department of Medicine, Division of Clinical Pharmacology, 2Department of Oncology; 3Department Background: Influenza virus infection is known to be of Surgery, 4Department of Physiology and basically self-limiting; however, severe condition, such Pharmacology, University of Western Ontario, as respiratory failure and encephalopathy, can London, Ontario, Canada sometimes occur especially in high-risk group of Corresponding Author: [email protected] patients, including children, and thorough treatments Conflict of Interest: None declared are required. Peramivir (Rapiacta®, Shionogi & Co., Ltd. / BioCryst), a new neuraminidase inhibitor, Background: Tamoxifen, a standard endocrine therapy administering intravenously, was approved for adults for estrogen receptor positive breast cancer, requires and a clinical trial for children was conducted in Japan hepatic metabolism by cytochrome P450 2D6 during the 2009-2010 influenza season. The data from (CYP2D6) to be converted to its active metabolite, the trials showed that peramivir is effective and safe endoxifen. Genetic variability and drug interactions can enough to use in both adults and children with cause alterations in CYP2D6 activity. Studies indicate influenza virus infection. that plasma levels of endoxifen are significantly lower Objectives: Because it is highly effective and the time in CYP2D6 poor metabolizers (PMs) and that PMs to recover from the infection is shortened after the have worse outcomes including recurrence rates and administration of mostly one dose, it was uncertain survival compared to EMs. However, recently there has e352 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 been conflicting evidence regarding the usefulness of codeine dose depending on her level of pain and CYP2D6 genotyping for tamoxifen therapy. CYP2D6 genotype. Objectives/Methods: We hypothesize that a patient’s Objective: To model the pharmacodynamic effect of CYP2D6 genotype and plasma endoxifen level codeine on pain levels in CYP2D6 retrospectively- together will better predict their success on tamoxifen. genotyped women recovering from c-section. We are collaborating with the London Regional Cancer Methods: Forty-five codeine-prescribed mothers Program to refer patients on tamoxifen to provide a provided a blood sample for CYP2D6 genotyping and blood sample for CYP2D6 genotyping by TaqMan recorded their pain level 4x/day for 3 days immediately assays and drug level analysis of tamoxifen and following a c-section. Genotyping was completed once endoxifen by LC-MS/MS. mothers had been discharged from hospital. Codeine Results: We observe that PMs have the highest doses and times were recorded, retrospectively adjusted tamoxifen plasma concentration compared to and modeled using the CYP2D6 genotype activity intermediate (IMs) and extensive metabolizers (EMs). score developed by Gaedigk et al and the Endoxifen levels for IMs and PMs are significantly pharmacokinetic data from Kirchheiner et al. lower than EMs. We observe a 12-fold variability in Results: No correlation (n=45, spearman’s rho= 0.034, endoxifen levels among EM patients, suggesting that p=0.827) was found between Area under the VAS-time other factors in addition to CYP2D6 contribute to Curve (AUC) for pain and genotype-adjusted codeine endoxifen concentration. We are examining the effects dose for the group. However, women at the genetic of interacting medications by drug level monitoring extremes reported codeine effects consistent with pre- and post-drug change and dose escalation of previous literature. The 2 PMs of codeine reported no tamoxifen in patients within the lowest quartile of analgesia as a result of taking codeine, while two of the endoxifen levels. three UMs reported immediate pain relief from Conclusions: Low endoxifen levels irrespective of codeine, but stopped taking it due to adverse affects CYP2D6 genotype may indicate a lack of benefit of (i.e. nausea, lightheadedness and constipation). tamoxifen therapy and may be a better predictor of Conclusion: A model to predict the analgesia affect of therapeutic outcome. codeine is more complex than its CYP2D6 genotype score alone. 80 Pharmacogenetics of post partum management 81 - a single center experience Pharmacogenomic prediction of anthracycline- vanderVaart1,2, Berger H3, Koren G1,2,4 induced cardiotoxicity in children 1Department of Pharmaceutical Sciences, University of Visscher H1, Ross CJD1, Rassekh SR2, Sistonen Toronto, Toronto, Ontario, Canada; 2Division of J1, Amstutz U7, Barhdadi A3, Dubé M-P3, al- Clinical Pharmacology and Toxicology, The Hospital Saloos H4, Sandor GS4, Brown AMK3,5, Rogers for Sick Children, Toronto, Ontario, Canada; 2 3,5 6 7 3 PC , Phillips MS , Rieder MJ , Carleton BC , Department of Obstetrics and Gynecology, St. 1 4 Hayden MR & the CPNDS Consortium Michael’s Hospital, Toronto, Ontario, Canada; Ivey 1Centre for Molecular Medicine and Therapeutics, Chair in Molecular Toxicology, Department of Child and Family Research Institute, Department of Medicine, University of Western Ontario, London, Medical Genetics, University of British Columbia, Ontario, Canada Vancouver, Canada; 2Division of Pediatric Corresponding Author: [email protected] Hematology/Oncology/BMT, Department of Pediatrics, Conflict of Interest: None declared University of British Columbia, Vancouver, Canada; 3Montréal Heart Institute Research Centre and Background: Codeine, a common opioid prescribed Université de Montréal, Montréal, Canada; 4Division of for pain post c-section, is biotransformed by the highly Pediatric Cardiology, Department of Pediatrics, polymorphic Cytochrome P450 enzyme 2D6 University of British Columbia, Vancouver, Canada; (CYP2D6). Ultrarapid metabolizers (UM), individuals 5Montréal Heart Institute and Genome Québec with multiple active copies of CYP2D6, can Pharmacogenomics Centre, Montréal, Canada; biotranform up to 50% more codeine into morphine 6Department of Paediatrics, Children’s Hospital at than normal, resulting in adverse reactions to codeine. London Health Sciences Centre, London, Canada; In contrast, poor metabolizers (PM), individuals who 7Pharmaceutical Outcomes and Policy Innovations have no active CYP2D6 genes, convert almost no Programme, Division of Translational Therapeutics, codeine into morphine and as a result may take Department of Pediatrics, University of British multiple doses of codeine without attaining analgesia. Columbia, Vancouver, Canada It would be optimal if we could titrate a mother’s

Corresponding Author: [email protected] 6Faculty of Medicine, University of Ottawa, Ottawa, Conflict of Interest: None declared Ontario; 7Health Canada, Ottawa, Ontario; 8Department of Laboratory Medicine and Pathology, Background: Anthracycline-induced cardiotoxicity Faculty of Medicine and Dentistry, University of (ACT) is a serious adverse drug reaction limiting its Alberta, Edmonton, Alberta; 9Department of Biology, use and causing substantial morbidity and mortality in Faculty of Science, University of Ottawa; 10Faculty of childhood cancer survivors. High cumulative doses are Medicine and Dentistry, University of Alberta, known to increase cardiotoxicity risk, but genetic Edmonton, Alberta; 11School of Pharmacy, Faculty of factors are suspected to be important as well given the Medical and Health Sciences, University of Auckland, high inter-individual variability in tolerated doses. Auckland, New Zealand; 12School of Pharmacy, Curtin Objective: Our aim was to identify genetic variants University of Technology, Perth, Australia; 13Research associated with ACT in patients treated for childhood Institute, McGill University Health Centre; cancer. 14Department of Pediatrics, and Department of Methods: We carried out a genetic association study Medicine, University of Alberta, Edmonton, Alberta; using 2977 single nucleotide polymorphisms (SNPs) in 15Women and Children’s Health Research Institute, 220 key drug biotransformation genes in a discovery Edmonton, Alberta cohort of 156 anthracycline-treated children from Corresponding Author: [email protected] British Columbia, with replication in a second cohort Conflict of Interest: None declared of 188 pediatric oncology patients from across Canada as part of the Canadian Pharmacogenomics Network Background: Many consumers use natural health for Drug Safety (CPNDS). products (NHPs) concurrently with prescription Results: We identified a highly significant association medications. As NHP-related harms are under-reported of a coding variant in a transporter gene with ACT through passive surveillance, the safety of concurrent (P=1.0x10-4). We found further evidence (P<0.01) for NHP-drug use remains unknown. associations with risk and protective variants in other Objective: To assess the feasibility of active genes including several other transporters. Combining surveillance in participating community pharmacies to these variants with important clinical risk factors in a identify adverse events related to concurrent NHP- predictive model, we classified patients into three risk prescription drug use. groups. In the high-risk group, 75% of patients were Methods: Participating pharmacists asked individuals accurately predicted to develop ACT, with 36% picking up prescription medications about 1) developing ACT within the first year; whereas in the concurrent NHP/drug use in the previous three months low-risk group, 96% of patients were accurately and 2) the presence of potential adverse events. If a predicted not to develop ACT. potential adverse event was identified and the patient Conclusions: We have identified multiple genetic agreed, a research pharmacist conducted a guided variants associated with ACT. Combined with clinical telephone interview to gather additional information. risk factors, genetic risk profiling can be used to Results: Over a total of 112 pharmacy weeks, 2615 identify high-risk patients who can then be provided patients were screened, of which 1037 (39.7%; 95% with safer treatment options. CI: 37.8% to 41.5%) reported concurrent NHP and prescription medication use. A total of 77 patients 82 reported a possible AE (2.94%; 95% CI: 2.4% to Study of Natural Health Product Adverse 3.7%), which represents 7.4% of those using NHPs and Reactions (SONAR): piloting an active prescription medications concurrently (95%CI: 6.0% to 9.2%). surveillance model in community pharmacies Conclusion: Compared to passive surveillance, this Vohra S1,2,3, Cvijovic K4,5, Boon H4, Foster B6,7, 5 8 8 7 study found active surveillance to markedly improve Jaeger W , LeGatt D , Cembrowski G , Murty M , NHP adverse event reporting rates. Active surveillance 7 7 2,9,10 11 Vu D , Leitch R , Tsuyuki R , Barnes J , is feasible and offers improved quantity and quality of Charrois T12, Arnason J9, Ware M13, Rosychuk R14,15 adverse event data, allowing for meaningful 1CARE Program, Department of Pediatrics, Faculty of adjudication to assess potential harms. Medicine, University of Alberta, Edmonton, Alberta; 2 School of Public Health, University of Alberta, 83 Edmonton, Alberta; 3Department of Medicine, 4 Adherence measurement methods among HIV University of Alberta, Edmonton, Alberta; Leslie Dan positive adolescents in Uganda: a prospective Faculty of Pharmacy, University of Toronto, Toronto, cohort pilot study Ontario; 5Department of Clinical Pharmacy and Diagnostics, University of Vienna, Vienna, Austria; e354 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

Wiens MO1,2, MacLeod S1,2, Musiime V3, overestimate adherence measured electronically. Ssenyonga M3, Kizza R4, Kitaka S5, Adome RO5, However, overall adherence measured with all methods Ssali F3 was still clinically acceptable. 1University of British Columbia, Vancouver, BC; 2Child and Family Research Institute, Vancouver, BC; 84 3Joint Clinical Research Centre, Kampala, Uganda; Determinants of CYP3A4 expression and 4Norra Stockholms Psykiatri, St Görans Hospital, activity in the Huh7 human hepatoma cell 5 Stockholm, Sweden; Makerere University, Kampala, model of non-alcoholic fatty liver disease Uganda Woolsey SJ1, Beaton MD3, Tirona RG1,2 Corresponding Author: [email protected] 1Department of Physiology and Pharmacology; Conflict of Interest: None declared 2Department of Medicine, Division of Clinical Pharmacology, 3Division of Gastroenterology, Lawson Introduction: The quality of traditional adherence Health Research Institute, The University of Western measurements among adolescents is difficult to assess. Ontario, London, Ontario, Canada Antiretroviral (ARV) adherence research among Corresponding Author: [email protected] adolescents living with HIV in resource-constrained Conflict of Interest: None declared countries is limited. Objectives: Our primary objective was determining Background: The recent rise in rates of obesity, feasibility of a large-scale, long-term study using diabetes and metabolic syndrome among the Western electronic adherence monitoring in Uganda. Our population, has caused a parallel increase in the secondary objective was to compare accuracy of pill- prevalence of non-alcoholic fatty liver disease count (PC) and self-report (SR) adherence with (NAFLD). A defining characteristic of NAFLD is electronic medication vials (eCAPs). eCAP’s record steatosis within hepatocytes. Despite the high compliance in real-time with data downloaded during incidence, little is known regarding the effect of refills. NAFLD on hepatic drug metabolism and its impact on Methods: Adolescents receiving care at the Joint drug response. We hypothesize that hepatic CYP3A Clinical Research Centre in Kampala, Uganda were activity is altered as a result of fat accumulation, recruited. ARV’s were dispensed in eCAPs for 1 year. leading to changes in the pharmacokinetics of everyday Person-pill-days (one day where adherence was drugs. measured for one medication in one person) were Objective: Establish an in vitro model of human calculated for each patient and a weighted paired t-test NAFLD to study regulation of expression and activity was used to compare the levels of adherence among all of CYP3A4. subjects for three different adherence methods. Methods: To better understand the mechanisms Results: Fifteen patients were included: 40% were involved in the regulation of CYP3A4 in NAFLD, we female, mean age was 14, mean baseline CD4 count will use the Huh7 human hepatoma cell line known to was 244, and average treatment duration was 9 months natively express CYP3A4. In the cell model, Huh7 at study entry. 4721 person-pill-days were observed. cells are incubated with free fatty acids to induce fat Several eCAPs required replacement during the study overloading similar to that found in steatotic liver. resulting in some data loss. Consent rate was high Lipid quantification, cytotoxicity and CYP3A4 mRNA (94%) but was slow due to age limit cut-points, and protein expression will be examined. Furthermore, indicating that a future study should be multi-site. A biotransformation activity will be assessed by longitudinal examination of the eCAP data showed that measuring the metabolism of midazolam, a probe drug while most non-adherence among individual subjects substrate for CYP3A4 enzymes. was time-dependant (during times of poor adherence Results: Preliminary data from our laboratory indicates all medications were affected), some was drug- that patients with NAFLD have altered CYP3A4 dependant (adherent to one medication but not others). expression and activity. Our small pilot sample precluded a regression analysis Conclusions: It is expected that this in vitro model of to further explore this effect. Overall compliance for human NAFLD will provide the platform for further SR was 99%, PC was 97% and eCAP was 88% studies aimed to define the signaling pathways (p<0.05 for all comparisons). 93%, 67% and 23% of involved in regulating CYP3A4 activity. Together with patients had a compliance of greater than 95% among studies in patients, these studies may provide a basis SR, PC and eCAP methods, respectively. for better and safer pharmacotherapy in NAFLD. Conclusions: A large-scale adherence study in Uganda is feasible using a more robust electronic monitoring system. Adherence measurements produced by pill counts and self reporting methods appear to

85 Conclusions: Our results demonstrated that H2S Hydrogen sulfide mediates the inhibitory effect mediates the inhibitory effect of sulforaphane on the of sulforaphane on the proliferation of human proliferation of PC-3 cells, which suggests that H2S- prostate cancer cells releasing diet or drug might be beneficial in the Guangdong Yang1, Yanxi Pei2, Qiuhui Cao3, treatment of prostate cancer. Lingyun Wu4, Rui Wang3 1School of Kinesiology, Lakehead University, Thunder Bay, Canada; 2College of Life Science, Shanxi University, Taiyuan, China; 3Department of Biology, Lakehead University, Thunder Bay, Canada; 4Department of Pharmacology, University of Saskatchewan, Saskatoon, Canada Corresponding Author: [email protected] Conflict of Interest: None declared

Background: Hydrogen sulfide (H2S) is a novel gasotransmitter that regulate cell proliferation and other cellular functions. Sulforaphane is a sulphur- containing compound that exhibits anticancer properties, and young sprouts of broccoli are particularly rich in sulforaphane. There is consistent epidemiological evidence that the consumption of sulphur-containing vegetables, such as garlic and cruciferous vegetables, may help reduce the occurrence of prostate cancer. Objectives: We want to determine whether H2S mediates the anti-survival effect of sulforaphane on prostate cancer and the underlying mechanisms. Methods: H2S level, cell viability, and protein expression were measured by methylene blue method, MTT assay, and western blotting, respectively. Results: A large amount of H2S is released from sulforaphane when sulforaphane was injected into mice or added into the cell culture medium or mixed with mouse liver homogenates. Both sulforaphane and NaHS (a H2S donor) decreased the viability of PC-3 cells (a human prostate cancer cell line) in a dose- dependent manner, and supplement of methemoglobin or oxidized glutathione (two H2S scavengers) reversed sulforaphane-reduced cell viability. NaHS also significantly inhibited PC-3 cell migration. We further found both cystathionine gamma-lyase (CSE) and cystathionine beta-synthase are expressed in PC-3 cells and mouse prostate tissues. H2S production in prostate tissues from CSE knockout mice was only 20% of that from wild-type mice, suggesting CSE is a major H2S- producing enzyme in prostate. CSE overexpression enhanced H2S production and inhibited cell viability in PC-3 cells. In addition, sulforaphane and NaHS activated p38 mitogen-activated protein kinases (MAPK) and c-Jun N-terminal kinase (JNK). Pre- treatment of PC-3 cells with methemoglobin decreased sulforaphane-stimulated MAPK activities. Suppression of both p38 MAPK and JNK reversed H2S- or sulforaphane-reduced viability of PC-3 cells. e356 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

AUTHOR INDEX

Brackstone M 79 Author Abstract No. Braun J 41 Acuna P 57 Brien J 8 Adams-Webber T 65 Brojeni PY 21 Adome RO 83 Brown AMK 81 Al-Aghawani W 19, 47 Burger D 50 Alam MS 48 Burland LB 39 Aleksa K 51, 59, 73, 74 CaccameseS 71 Aljudaibi B 9 Cao Q 85 AL-Lahham A 19 Carey N 22 Al-Madi S 19 Carleton B 23 Almazan G 56 Carleton BC 6, 35, 41, 69, 81 al-Saloos H 81 Carnevale A 51 Amstutz U 41, 81 Carvalho JC 35 Arburg B 39 Casper RF 24 Armstrong C 20, 34, 70 Castaneda-HernandezG 23 Arnason J 82 Castelan-Martinez OD 23 Balicki D 20 Cembrowski G 82 Barhdadi A 81 ChandeN 9,76 Barnes J 82 Charbel C 52 Barrera M 75 Charrois T 82 Bawolak MT 72 ChatenetD 53 Beaton MD 84 Chaudhry S 10, 54 Beauchemin S 7 ChemtobS 30,43,49 Beauchemin S 13 Chen N 60 Beauclair L 25 ChiassonJL 31 Bélanger F 20, 31, 34, 70 Chouinard G 55 Bend JR 33 Chu S 1 Berger H 10, 80 Chuang AI 11 Bhosle V 30 CiszkowskiC 6 Blais M 49 Clark CA 24 Blanchette A 52 Cloos JM 55 Bocking A 8 Côté J 58 Bocquet V 55 CPNDS consortium 81 Boon H 29, 82 Crépeau J 67 Bosma KJ 38 CvijovicK 82 Bournissen FG 21 DallaireS 31 Bovenzi V 58 DavidovitsJ 24

De Paula M 56 Gorodischer R 12 de Wildt SN 28 Gregor JC 9, 76 DeGorter MK 2, 42 Gunness P 59 DesmaraisJE 25 Ha EH 61 Dingle B 79 Ha M 61 Doucet J 31 Hackmon R 32 Dresser GK 9, 38, 76 Hamel D 30 Dubé MP 81 Hamet P 31 Dubuc C 58 Han JY 61 Dupuis J 53 Hanly L 60 Eichhorst J 32 Harding R 8 Elzagallaai AA 3 Hardy P 43 Feng Q 46 Harper DP 11 Ferguson KA 40 HaydenMR 6,35,41,69,81 Fierro-Evans M 23 Hegmann T 16 FilippelliA 29 Hong SC 61 FinkelsteinY 26,57 Hong YC 61 FlockhartDA 71 Hou X 30 Fortin D 58 Huguet J 31 Foster B 82 HutsonJR 5,26,32 Fournier A 53 IglesiasL 31 Fu A 18 ItoS 11,27,57 Fujii H 27 Jaeger W 82 Gaedigk A 27 JahedmotlaghZ 33 Gagnon C 43 Jayyab AA 19, 47 Garcia-Bournissen F 5, 27, 57 Jimenez-Mendez R 23 Garcia-Rojas Y 23 Jizi K 62 Gardiner P 29 Jong GW 63 GaudetteF 20,31,70 Juneau M 67 George M 57 Juurlink DN 26 Gera L 72 Kapur BM 15, 32 Gijsen VMGJ 28 Kelly L 6, 64 Gillis AM 39 Kenna K 8 Giraldeau G 67 Kennedy DA 65 2, 4, 9, 14, 38, 42, 66, Girard L 29 Kim RB 76, 79 Girdwood AJ 66 Kim Y 61 Gladdy R 35 Kitaka S 83 GobeilFjr 58 Kizza R 83 Gong IY 4 Knauer MJ 42, 66 e358 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

Koch P 55 MatokI 12,15,21 3, 5, 6, 8, 10, 12, 15, MichaudJ 7,13 21, 22, 24, 26, 28, 32, 33, 35, 37, 40, 51, 54, MillerDW 16,36 57, 59, 60, 61, 63, 64, Koren G 65, 67, 73, 74, 80 MiyazakiH 44 Kost S 45 Morgan AD 14 Lam J 6 Murty M 82 Lam K 26 Musiime V 83 Laskin CA 24 Nagai Y 78 Lausman A 10 Natekar A 74 LeblondFA 7,13 Naud J 7, 13 Ledoux J 52 Nava-Ocampo AA 61 Lee CYW 67 Nelson M 31 Leeder S 27 Nguyen QT 53 Lefrancois S 7 NulmanI 54,75 LeGatt D 82 On N 36 Leitch R 82 Orbach H 12 Lepage M 58 Osadchy A 37 LétourneauM 53 Ovakim DH 38 Leung YH 34 Park H 61 Levine MAH 68 Parkinson FE 1, 45 Levy A 12 Patel V 76 Liu M 46 Pei Y 85 Liu Y 46 Phillips MS 81 Loo TT 69 Picard E 43 Lu J 70 Pichette V 7, 13 Lu WJ 71 Pollak PT 39, 77 Lu X 46 Ponich T 9, 76 LubetskyA 5,32 Pope E 57 Luo S 18 Rassekh SR 41, 69, 81 MacLeod S 83 RatnapalanT 37 Madaan A 49 RiddickDS 17 MadadiP 6,10,35,64 RiederMJ 3,40,60,64,81 Maghni K 62 Rivas-Ruiz R 23 Mansell SE 4 Rivera JC 30, 49 Marceau F 72 Rogers PC 81 Marcotte F 67 Rolland-Portal I 55 Margolese HC 25 Ross C 41, 64 Marques-Avila S 23 RossCJD 35,69,81 Matlow J 73 RosychukR 82

RotenbergB 40 Tsuyuki R 82 Russell E 40 Turgeon J 20, 31, 34, 70 Sakaguchi S 27 Urquhart BL 2, 9, 18 Sanchez M 30 Utgikar R 17 Sandor GS 81 Uum SV 40 Sapieha M 49 Vandenberg T 79 Sarkar M 65 vanderVaart S 80 Savard M 58 Veenstra F 64 Schoeman K 33 Velenosi TJ 18 Schwarz UI 9, 76 Villeneuve LR 52 Sen M 38 Visscher H 81 Shannon MW 57 Vohra S 29, 82 Shaw K 41 Vu D 82 Shear NH 57 Walfisch A 5 Sheiner E 12 Walker D 8 Siddiqui N 48 Wang H 18 Silverman G 35 Wang R 85 Sistonen J 35, 64, 81 Ware M 82 Sivilotti MLA 26 Welman M 62 Soon GS 57 Wiens MO 83 Spitzer KA 24 Wiznitzer A 12 Ssali F 83 Woolsey SJ 84 Ssenyonga M 83 Wu L 85 Stables JP 48 Wyse DG 39 Stern SJ 15, 65 Xiang FL 46 Sugiyama D 42 Xiong W 45 Sun C 45 Yang C 43 Sun Z 16 Yang G 85 Syed U 68 Yang MD 11 Tahiri H 43 Yathindranath V 16 TakitaniK 44 Yeger H 60 Tamai H 44 Young GB 38 Tanaka T 78 Younus J 79 Tardif JC 67 Zelner I 8 TeftWA 42,79 ThompsonCS 50 2, 14, 38, 42, 66, 76, Tirona RG 84 ToussaintF 52 Touyz RM 50 e360 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

KEY WORDS INDEX

Key Words Abstract No. Children 28, 51, 57 6-mercaptopurine 5 Choroid: neovascularization 43 ABC transporters 7 Cisplatin 23 2 Abuse 55 Clinical equivalence 5 Active surveillance 82 CNS depression 10 Acyclovir 59 Cocaethylene 74 Adenosine 1, 45 Codeine 6,10,64,80 Adenovirus 42 Colorectal cancer 65 Adherence 83 Convolvulus arvensis 19, 47 Adolescents 83 Cortisol 40 Adverse drug reaction 3, 23, 35 Crohns disease 9, 76 Amiodarone 39, 77 CYP2C19 27 Angiogenesis 49 CYP2D6 79 Antagonist 53 CYP3A4 84 Antenatal exposure 37 CYP450s 20 Anthracyclines 81 Cytochrome P450 13, 17 Anticonvulsants 48 Cytochrome P450 2D6 (CYP2D6) 80 Antimalarials 37 Cytochrome P450 3A 11 Antiviral drug 78 Dehydration 11 Aortic rings 19 Dehydroandrosterone 44 Apnea-hypopnea index 40 Drug excretion 4 Azathioprine 9, 75 Drug hypersensitivity 3 B2 receptor 72 Drug metabolism 17, 84 BCRP 59 Drug safety 12 Behaviour 45 Drug toxicity 34 Benzodiazepine 55 Drug transport 2, 7, 31 Biomarker 74 Drug use review (DUR) 68 Birth Defects 12 Drug-drug interactions 31, 70 Blood pressure 53 Drugs 29 Blood-brain barrier 36, 58 Endothelial 50 Bradykinin 72 Endothelial cells 52 Breast cancer 20, 79 ENT1 45 Breastfeeding 6, 22 Epicardium derived cells 46 CaMKII 52 Epidemiology 26 Cancer 22,58 Ethanolteratogenicity 8 Cardiotoxicity 81 Ethics 54 Cellaccumulation 16 Ethyl glucuronide 73

Excitotoxicity 1 Lacticacid 34 Fattyacidethylesters 8 Lysophosphatidic acid (LPA) 36 Fetal alcohol spectrum Maximakinin 72 disorder 73 Meconium 8 Fetalsafety 21 Metabolism 20 Fishconsumption 33 Methanol 32 Folate 61 Methods 68 Folicacid 15,65 Methotrexate 76 Folicacidsupplement 61 Methylmercury 33 Formic acid 32 Microparticles 50 GC/MS 51 Monitoring 39 Genericsubstitution 25 MTHFR C677T Genotype 31 polymorphism 65 Glibenclamide 19 Myocardial infarction 46 Growth factor receptors 43 N-acetylcysteine 60 Growth factors 43 Neonates 10 H2S 85 Neovascularization 43 Hair 74 Nephrotoxicity 28, 60 Hairbiomonitoring 33 Neuraminidaseinhibitor 78 hENT1 1 Neuropathy 69 Herb-drug interactions 29, 82 Neurotoxicity 48 Herbs 29 NewZealandrabbit 19 HIV 83 NFAT5/TonEBP 11 HNF4alpha 66 NF-kB 13 Human prostate cancer cells 85 NK1 receptor antagonist 62 Non-alcoholic fatty liver Hypertension 52 disease 84 Hyperthyroidism 77 Non-institutional 68 Ifosfamide 60 Nuclear receptors 66 Immunosuppressive drugs 62 OATP transporters 70 In vitro diagnosis 3 Oatp1b2knockoutmouse 2 In vitro to in vivo prediction 14 OATP2B1 66 Infant 27 Obesity 15 Inflammatory bowel disease (IBD) 75 Obstructivesleepapnea 40 Influenzavirusinfection 78 Oculartoxicity 37 Interferon alpha 21 Opioids 35 Intrinsic clearance 14 Ototoxicity 23 Ironoxidenanoparticle 16 Oxidative Stress 50 Isatin 48 Oxycodone 6 Paediatric obstructive sleep KCL 47 apnea 64 Kinin 58 e362 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

Paediatrics 41, 69 Stem cell factor 46 Pain 80 Stem cells 49 Pantoprazole 27 Stevens-Johnson Syndrome 57 PBDE hair analysis 51 Sulforaphane 85 Permeability 36 T cells 62 P-glycoprotein 4, 14 Tacrolimus 28 Pharmacogenetics 35, 41 Tamoxifen 79 Pharmacogenomics 69, 81 Teratogen exposure 54 Pharmacokinetic model 14 Teratogenic 24 Pharmacokinetics 15, 76 Toxic epidermal necrolysis 57 Pharmacy error 77 Toxicity 16,39 Phenylephrine 19 Toxicology 26 Placenta 5, 32 Transporter 42 Placental perfusion 73 Transporters 34, 66 Platelets 3 Ulcerative colitis 9 Poisoning 26 Urotensinergicsystem 53 Polycyclic aromatic VAS 80 hydrocarbons 17 Vincristine 69 Pravastatin 24 Warfarin 41 Prediction 42 Pregnancy 5, 12, 21, 22, 75 Pregnancy termination 54 Preterm delivery 61 Prevalence 55 Psychotropic medications 25 PXR 13 Rabbit'saorticrings 47 Recurrent pregnancy loss 24 Renal 59 Renal disease 7 Respiratory depression 64 Retina 49 Retinol 44 Retinolmetabolic enzyme 44 Rivaroxaban 4 Safety 82 Solute carriers 7 Statin 70 Statinpharmacokinetics 2 Steatosis 84