|Bioluminescent Approaches for In Vitro ADMET|
James J Cali, Mary Sobol, Dongping Ma, Michael Valley
Bioluminescent ADMET assays couple targets of interest to the photon emitting reaction of firefly luciferase. Multiplexed cell-based assays measure compound cytotoxicity and induction of P450 transcription and enzyme activity. Cell-free membrane assays measure P450 or monoamine oxidase enzyme activities and their inhibition by test compounds.
|Initiatives For The Sharing Of Toxicity Data|
Much high-quality toxicity information is held within organisations rather than being in the public domain. Organisations would benefit from sharing this data with each other in a reciprocal manner where commercial sensitivity issues allow.
|MS-Xelerator™: Advanced Algorithms for LC/MS Data Processing Applied to Biomarker Discovery, Differential Analysis and Quantitative Proteomics|
LC-MS based proteomic experiments are used to compare complex biological samples across multiple conditions. Fast, powerful computational tools are needed to explore and detect differences in the areas of Expression Proteomics and Biomarker Discovery. In general, specialized steps are necessary to solve these difficult problems (binning, alignment & normalization, peak picking, relative quantitation, etc.). MS-Xelerator is a collection of software tools dedicated to all of the above tasks.
|A Robust High Throughput Physicochemical Screen for Phospholipidosis|
Pavol Vitovic, Juha-Matti Alakoskela and Paavo K.J. Kinnunen
The poster presents a simple HT physicochemical screen, predicting PLD in real-time by surface activity profiling (using Kibron Delta-8 analyzer). The results are equal or better than derived from a cell based assay, throughput is up 450 compounds/24 hrs and drug consumption < 1 mg/compound.
|Promega’s Multiplexed Cell Viability and Apoptosis Assays Performed on the PHERAstar|
Tracy Worzella and Brad Larson
Today’s high-throughput screening facilities face increasing demands to generate more information from their existing compound libraries. In this poster, we demonstrate the combination of several Promega cell-based assays (CellTiter-Blue®, Apo-One® and Caspase-Glo® 3/7) multiplexed in both low-volume 384 and 1536-well plate formats. The BMG LABTECH PHERAstar microplate reader is used to record both luminescence and fluorescence, depending on the multiplex combination.
|Promega’s Multiplexed Luciferase Reporter and Cell Viability Assays performed on the PHERAstar|
Tracy Worzella and Brad Larson
By multiplexing a reporter assay (EnduRen™, ViviRen™) with a cell viability assay (CellTiter-Glo®), it is possible to determine if reporter response variations are due to changes in cell number and health. In this poster, we demonstrate the combination of several Promega cell-based assays multiplexed in both low-volume 384- and 1536-well plate formats using the BMG LABTECH PHERAstar microplate reader to record luminescence.
|Metabolic Stability and Clearance of Pharmaceutical Chemicals in Pre-Pooled Cryopreserved Hepatocytes|
Aruna Koganti, Nicola J. Hewitt, Wei Zhang, Michael Cheseborough, Christopher M. Terrell, Paul M. Silber and Charles B. Jensen
Cryopreserved hepatocytes express both phase I and phase II enzymes and facilitate early evaluation of the metabolic stability of pharmaceuticals. Availability of pre-pooled cryopreserved hepatocytes further enhances the utility of this model by reducing donor to donor variability. In this study, the metabolic stability of 29 pharmaceutical compounds were evaluated in pre-pooled cryopreserved human hepatocytes.
|CYP4F Enzymes are the Major Enzymes in Human Liver Microsomes that Catalyze the O-Demethylation of the Antiparasitic Prodrug DB289|
Greg Loewen, Michael Zhuo Wang, Janelle Saulter, Etsuko Usuki, Yen-Ling Cheung, Michael Hall, Arlene Bridges, Oliver Parkinson, Chad Stephens, James Allen, Darryl Zeldin, David Boykin, Richard Tidwell, Mary Paine, James Hall and Andrew Parkinson
DB289 is a prodrug that is converted by several steps to the active metabolite DB75. DB289 exhibits enhanced oral efficacy and reduced acute toxicity over DB75, an aromatic dicationic compound that is effective against a broad range of pathogens in vitro including African trypanosomiasis (African sleeping sickness). This reaction phenotyping study aimed to identify the enzymes responsible for oxidative O-demethylation; the first step in this conversion to DB75.
|Effects of Gender, age and Ethnicity on Human Cytochrome P450 Activity|
Lisa Collins, L. Anne Dwyer, Zell Woodworth, Tiffin Ramsey, Clayton Otwell, Chad Pope, Stephanie Helmstetter, Terry Graves, Josh Snyder, Jason Barricklow and Andrew Parkinson
The expression of CYP enzymes is influenced by both endogenous and exogenous factors. The aim of this analysis was to evaluate whether the age, gender, or ethnicity of the donor should influence the selection of human liver microsomes for drug metabolism studies, as well as whether cigarette smoking and alcohol consumption are reliable indicators of elevated CYP1A2 and CYP2E1 activity, respectively.