2^nd International Conference on Clinical Trials and Therapeutic Drug Monitoring August 22-24, 2016 Philadelphia, Pennsylvania, USA

Anna Bejrowska has graduated from Gdansk University of Technology (Poland) in the year 2013. She is a co-author of a paper about environmental tests based on nuclear receptors’ activity changes. Her curiosity and desire for intellectual development is what led her to pursue her PhD in the field of Drug Development. Currently, she is a PhD candidate in the Department of Pharmaceutical Technology and Biochemistry at Gdansk University of Technology. She is consistently developing her experience of working with active compounds, enzyme fractions and cell cultures. Her interest is focused on modulations and differentiation of drugs’ metabolism, including the role of nuclear receptors in those processes.

Modern cancer treatment provides promising outcomes, thanks to the combined therapies. However, aiming to decrease resistance against individual drugs, treatment can intensify their adverse and toxic effects. Therefore, it is important to examine the influence that potential therapeutics have on cellular metabolism. Our group previously revealed that antitumor acridinone derivatives, C-1305 and C-1311, are metabolized to a great extent by UGT1A10. Thus, the aim of the present study is to test the ability of these compounds to modulate the activity of UGT1A10 isoenzyme both in non-cellular and cellular systems. The experiments were performed using human recombinant isoenzyme UGT1A10 and colon cancer line HCT-116 over expressing UGT1A10. Enzyme activity in both models were measured using UGT-specific reaction, 7-hydroxy-4-(trifluoromethyl)-coumarin glucuronidation in the presence of selected acridinone derivatives, as well as without the drug (control experiments) by RP-HPLC analysis. The results showed that, C-1305 and C-1311 act differently towards UGT1A10 activity in dependence on the applied model. Enzymatic activity of UGT1A10 was reduced by both acridinone derivatives in non-cellular system. By contrast, higher level of UGT1A10 activity was observed in HCT-116 cells treated with both studied compounds. It is supposed that C-1305 and C-1311 potentially applied in multidrug therapy might modulate the effectiveness of UGT1A10 on the protein and the transcriptional level. This finding provides new insights into potential pharmacokinetic drug-drug interactions between C-1305 and C-1311 and the substrates of UGT1A10.

Anna Mróz has studied Biotechnology from Gdańsk University of Technology (Poland) and received her Master’s degree in the year 2013. She is now a PhD student at the Department of Pharmaceutical Technology and Biochemistry, Gdańsk University of Technology. Her scientific interests are related to the investigation of metabolism of new active compounds against cancer. She is a co-author of two papers in the field of UDP-glucuronosyltransferases-mediated metabolism.

UDP-glucuronosyltransferases (UGTs) are phase II conjugating enzymes catalyzing glucuronidation reaction of many exogenous and endogenous substances. Glucuronides are more polar than the native compounds which facilitates their excretion. UGT isoenzymes are in the spotlight in respect to antitumor therapy as glucuronidation is the major route of elimination for many anticancer drugs. Moreover, some drug-drug interactions can be explained on the basis of UGTs inhibition. The objective of the present study was to explore the inhibitory effect of C-1748, an acridine antitumor agent, toward isoenzyme UGT1A9 in order to predict the potential for drug-drug interactions. C-1748 showed strong cytotoxic activity against colon cancer cells and high anti-tumor activity against prostate carcinoma xenografts which allowed its selection for preclinical studies. Previous results indicated that C-1748 undergoes glucuronidation with human liver and intestinal microsomes. Our studies showed that C-1748 is not a substrate for UGT1A9, however it exerted dose-dependent inhibition toward this isoenzyme. The glucuronidation reactions of standard substrate 7-hydroxy-4-(trifluoromethyl)coumarin were conducted in the absence or presence of C-1748 and monitored by HPLC/UV-Vis method. The concentration of 0.25 mM of C-1748 inhibited more than 70% activity of UGT1A9. The inhibition kinetic type was determined to be noncompetitive on the basis of Lineweaver-Burk and Dixon plots and the inhibition kinetic parameter (Ki) was calculated to be 0.17 mM. Taking together, the presented UGT1A9 significant inhibition indicates the high possibility for drug-drug interactions in combined therapies using C-1748 anticancer agent. Therefore, clinical monitoring should be applied when co-administrating C-1748 with drugs mainly undergoing UGT1A9-mediated metabolism.

Javier del Pino has received his PharmD degree from the University Complutense University of Madrid in the year 2004. He has done two Master’s in Sciences in the year 2009 and 2010. He did his Specialization in Neurotoxicology and Neurodevelopmental Toxicology and received his PhD in Toxicology in the year 2009. In 2010, he worked at Institute of Health Carlos III in the National Center of Environmental Health. From 2010 to 2012, he was an Associated Researcher at University of Massachusetts (UMASS), working at Sandra Petersen´s Lab in a National Institute of Health (NIH) project on developmental effects of TCDD endocrine disruptor on sexual differentiation. In 2016, he became an Associate Professor of Toxicology at the Complutense University of Madrid, Spain.

Chlorpyrifos (CPF) is an organophosphate insecticide reported to induce both after acute and repeated exposure learning and memory dysfunctions, although the mechanism is not completely known. CPF produces basal forebrain cholinergic neuronal loss, involved on learning and memory regulation, which could be the cause of such cognitive disorders. This effect was reported to be mediated through apoptotic process, although neuronal necrosis was also described after CPF exposure. Accordingly, we hypothesized that CPF induces basal forebrain cholinergic necrotic and apoptotic cell death. We evaluated in septal SN56 basal forebrain cholinergic neurons, the CPF effect after 24 h and 14 days exposure on the necrosis induction and the apoptotic and necrotic gene expression pathways. This study shows that CPF induces after acute and long-term exposure necrotic cell death at higher concentrations than which induces apoptotic cell death. Evaluation of cell death pathways revealed that some of them are altered at lower concentrations than which produces the effects observed and below the no observed adverse effect level (NOAEL). The present finding suggests that the use of gene expression profile could be a more sensitive and accurate way to determine the NOAEL.

Javier del Pino has received his PharmD degree from the University Complutense University of Madrid in the year 2004. He has done two Master’s in Sciences in the year 2009 and 2010. He did his Specialization in Neurotoxicology and Neurodevelopmental Toxicology and received his PhD in Toxicology in the year 2009. In 2010, he worked at Institute of Health Carlos III in the National Center of Environmental Health. From 2010 to 2012, he was an Associated Researcher at University of Massachusetts (UMASS), working at Sandra Petersen´s Lab in a National Institute of Health (NIH) project on developmental effects of TCDD endocrine disruptor on sexual differentiation. In 2016, he became an Associate Professor of Toxicology at the Complutense University of Madrid, Spain.

Cadmium is a neurotoxin compound which induces cognitive alterations similar to those produced by Alzheimer’s disease (AD). However, the mechanism through which cadmium induces this effect remains unknown. In this regard, we described in a previous work that cadmium induces a more pronounced cell death on cholinergic neurons from basal forebrain. Degeneration of basal forebrain cholinergic neurons, as happens in AD, results in memory deficits attributable to the loss of cholinergic modulation of hippocampal synaptic circuits. Moreover, cadmium induces acetylcholinesterase (AChE) overexpression, which has been related to cell death induction. Moreover, AChE variants alteration has been reported to mediate apoptotic and necrotic cell loss induction of basal forebrain cholinergic neurons and development of AD. According to all above, we hypothesized that cadmium induces the more pronounced cell death on basal forebrain cholinergic neurons through alteration of AChE variants expression alteration. The present study is aimed at researching the mechanisms of cell death induced by cadmium on basal forebrain cholinergic neurons. For this purpose, we evaluated, in SN56 cholinergic murine septal cell line from basal forebrain region, the cadmium toxic effects on neuronal viability through AChE splice variants. This study proves that cadmium induces cell death on cholinergic neurons through overexpression of tetrameric AChE-S and down-regulation of monomeric AChE-R. Our present results provide new understanding of the mechanisms contributing to the harmful effects of cadmium on cholinergic neurons and suggest that cadmium could mediate this effect through AChE splices altered expression.

Javier del Pino has received his PharmD degree from the University Complutense University of Madrid in the year 2004. He has done two Master’s in Sciences in the year 2009 and 2010. He did his Specialization in Neurotoxicology and Neurodevelopmental Toxicology and received his PhD in Toxicology in the year 2009. In 2010, he worked at Institute of Health Carlos III in the National Center of Environmental Health. From 2010 to 2012, he was an Associated Researcher at University of Massachusetts (UMASS), working at Sandra Petersen´s Lab in a National Institute of Health (NIH) project on developmental effects of TCDD endocrine disruptor on sexual differentiation. In 2016, he became an Associate Professor of Toxicology at the Complutense University of Madrid, Spain

4-Aminopyridine (4-AP) is a potassium channel blocker used for the treatment of neuromuscular disorders. Otherwise, it has been described to produce a large number of adverse effects, among them cell death mediated mainly by blockage of K+ channels. Specifically, 4-AP has been reported to produce cell death in central nervous system on hippocampal cells. On the other hand, Kv channel interacting protein 1 (KChIP1) is a neuronal calcium sensor protein that is predominantly expressed at GABAergic synapses and it has been related with modulation of K+ channels, GABAergic transmission and cell death. According to this, KChIP1 could modulate K+ channels and GABAergic transmission, which mediate the toxic effects induced by 4-AP. We evaluated, in wild type and KChIP1 silenced primary hippocampal neurons, the effect of 4-AP (0.25 mM to 2 mM) with or without semicarbazide (0.3 M) co-treatment after 24 h and after 14 days 4-AP alone exposure on KChIP1 and Kv 4.3 potassium channels gene expression and GABAergic transmission. We observed that 4-AP modulates KChIP1 which regulates Kv 4.3 channels expression and GABAergic transmission. Our study suggests that KChIP1 is a key gene that may have a protective effect up to certain concentration after short-term 4-AP treatment, but this protection would be erased after long term exposure, due to KChIP1 down-regulation predisposing cell to 4-AP induced damages. These data might help to explain protective and toxic effects observed after overdose and long term exposure.

Sara Alzein has completed her PharmD and Bachelor’s degree of Pharmacy from Lebanese International University (LIU). She participated in the 7th Pharmacy Day “facts and myths in pharmacy practice” in May 2012 with a poster presentation, in LIU. The Abstract of her PharmD thesis entitled “pregnancy complications in the Lebanese population” which was accepted by the American Journal of Health-System Pharmacy (ASHP).

Pregnancy is associated with multiple health problems, which can be controlled by raising awareness about the complications that occur. Many Lebanese females in their childbearing age are not knowledgeable about the importance of folic acid intake and the risk of developing gestational diabetes mellitus (GDM) or urinary tract infections (UTI) during pregnancy. The aim of this study was to check whether females have been screened for GDM, counseled for UTI and the intake of periconceptional and postconceptional folic acid. The study was conducted in females that experienced at least 1 pregnancy in a community setting by filling a 10 minutes survey. Concerning GDM, 42.3% stated their knowledge of its screening importance, whereby 59.1% were screened with an OGTT and 32.8% with an FBG. This resulted in having 3.5% positive tests for this type of diabetes (3.1% had gained <18 kilos; p-value>0.05) for which 69% implemented dietary changes and 46% took metformin. As for folic acid intake, 85.6% took 5 mg dose: 33.1% for 1 month before gestation, 46.7% in the first trimester, 27.3% up to the second, and 11.5% throughout the pregnancy. It was noted that a big percentage of the females don’t know the role of GDM screening and its impact on pregnancy. Moreover, the role of periconceptional intake of folic acids is not sufficiently disseminated to young women, whereas the postconceptional was appropriate, but not based on risk factors. Furthermore, therapeutic management of UTIs in pregnancy requires thorough understanding of antimicrobial agents to optimize maternal and fetal outcomes.

Chung Yue Ling graduated from National University of Singapore in 2013 with a Bachelor of Science (Pharmacy) degree, First Class Honours. She is now working as an inpatient hospital pharmacist at the National University Hospital of Singapore, taking charge of general medicine and oncology wards.

This study aims to assess the adequacy and appropriateness of the use of chemical prophylaxis/enoxaparin in total knee and/or hip replacement (TKR and/or THR) surgery patients at National University Hospital (NUH). A retrospective drug utilization evaluation was performed for NUH patients aged ≥18 years old who have undergone TKR and/or THR surgery from 1st January to 31st March 2013 and excluded foreigners not residing in Singapore. The study indicators included compliance/non-compliance of chemoprophylaxis/enoxaparin prescribing patterns to NUH Venous-Thromboembolism (VTE) Prophylaxis guidelines. Efficacy and safety related clinical outcomes in terms of VTE and hemorrhagic events respectively in a 3-months follow-up period post surgery were also measured. Data for a total of 82 patients were collected and analysed. Chemoprophylaxis prescribing patterns for only 46 (56.1%) patients were compliant to NUH guidelines in terms of indication. The need for chemoprophylaxis exceeded bleeding risks for 55 (67.1%) patients but only 30 (36.6%) patients were given chemoprophylaxis (enoxaparin). When enoxaparin was prescribed, none of the dosing regimens were compliant to NUH guidelines in all three aspects of dose and frequency, prophylaxis duration and time of first dose initiation. During the 3-months follow-up, no bleeding events due to enoxaparin occurred. 9 (11.0%) patients developed thrombosis. Among which, one patient developed pulmonary embolism while another developed thrombosis in the femoral vein. The study revealed the baseline chemoprophylaxis and enoxaparin usage patterns in NUH TKR and THR patients. The adverse clinical outcomes that occurred identified potential safety gaps within the prescribing practices, for which recommendations were made to improve the safe and effective use of VTE chemoprophylaxis in NUH post-surgical orthopedic patients.