The Exploratory Pharmacology facility conducts a variety of industry-standard experiments to assess the drug-like properties of compounds including in vitro assays of compound stability, absorption, distribution, metabolism, excretion and toxicity (ADME/T). The core also supports the development of novel compounds for use in vivo applications by performing excipient screening, formulation studies, dose-ranging, tissue distribution studies, and comprehensive drug metabolism and pharmacokinetics (DMPK) studies. The core combines state-of the-art technology, including those listed below, and the expertise of academic and industry-savvy investigators to analyze and interpret the data to advance to most promising compounds.
In Vitro Analysis
- Aqueous Solubility
Compound stability in an aqueous solution is measured using an automated kinetic solubility method. The concentration of the compound in a saturated pH-buffered aqueous solution is determined by UV absorbance (250-498 nm) and compared to the spectra of a precipitation-free reference solution.
- Hepatic Microsome Stability
Metabolic stability of compounds is determined at a single concentration using species specific liver microsomes that contain many drug-metabolizing enzymes (including cytochrome P450s (CYPs), flavin monooxygenases, carboxylesterases, uridine glucuronide transferase and epoxide hydrolase). S9 fractions may be substituted for microsomes to assess the effects of additional metabolizing enzymes (aldehyde oxidase, glutathione transferase, monoamine oxidase, and sufurotransferase) on compounds metabolism.
- Plasma Protein Binding
The percentage of compound bound to plasma proteins is determined by rapid equilibrium dialysis and quantified by LC/MS/MS.
The potential toxic effects of compounds on cells are determined using a homogeneous, luminescence assay that measures the number of dead cells in culture using V79MZ cells (low metabolic activity) and primary or immortalized human hepatocytes (high metabolic activity).
- Cellular/BBB Permeability
Compound permeability is measured using an automated high-throughput, non-cell based, parallel artificial membrane permeability assay (PAMPA). The assay can be modified to predict trans-cellular intestinal absorption (gut permeability) or blood-brain-barrier diffusions (CNS penetration).
- Bioanalytical Method Development and Sample Analysis
Bioanalytical method development and sample analysis provides quantitative measurement of an active drug and/or its metabolites in biological matrices (plasma, brain tissue, urine etc.). Utilizing state-of-the-art LC/MS/MS and HPLC instrumentation, bioanalytical methods are developed and optimized for sensitivity, recovery, and reproducability to provide our clients with accurate, high quality bioanalytical data.
- PAMPA-based Excipient Screening
A quick, cost-effective, and reasonably accurate method, suitable for use in preclinical development, to assess the effect of excipients on the permeability of sparingly soluble drug candidates, and guide formulation efforts for use of the compound in vivo.
- CYP450 Inhibition Profiling
Compounds that inhibit P450s may cause the toxic accumulation of other substrates. Evaluation of the test compounds as inhibitors of selected human CYP450s (1A2, 2C9, 2D6, 3A4) is performed using isoenzyme specific P450-Glo™ assay kits (Promega).
- Plasma Stability
The stability of a compound in plasma is measured by LC/MS/MS.
In Vivo Drug Metabolism and Pharmacokinetic Assays
- Rapid Assessment of Compound Exposure
This experiment is a rapid and efficient compressed in vivo PK screening method used to determine an estimated in vivo exposure (AUC) of novel chemical probes. RACE can be used for dose finding studies, the assessment of formulations, and tissue distribution.
- Comprehensive Pharmacokinetic Analysis
Compounds that show promising in vitro drug like properties, or that are more advanced are subjected to a comprehensive pharmacokinetic analysis. Standard pharmacokinetic parameters including Cmax T1/2, bioavailability (%F), and AUC are determined.
- Results Reporting
A number of different reporting options are available from summary data to full written reports.
- TECAN Freedom Evo150 High-throughput/High-capacity system for automated liquid handling with an InfiniteM200 Plate reader for UV/Vis.
- AB-SCIEX API-Turbo 4000 MS/MS for the accurate quantification of compound by tandem mass spectrometry. The core is equipped with 2 systems, specifically configured for rapid high-throughput analysis of small molecule compounds.
- AB-SCIEX 2000 MS/MS is available for assays requiring less dynamic range than the API-Turbo 4000, and for compound QC by MS.
- Waters Aquity Ultra-High performance liquid chromatograph is coupled with the API-Turbo 4000 for the rapid analysis of samples by LC/MS/MS.
- The second AB-SCIEX 4000, and the 2000 are each coupled to a Shimadzu Nexera Ultra High-Performance Liquid Chromatograph.
- TopCount NXT™ Microplate Scintillation Counter is available for assay requiring radiolabeled tracers.
For a Price List, please call (407)745-2064 or email us.