Bioanalytical Chemistry, Drug Metabolism, and Pharmacokinetics

The BCDMPK Group at EIDD is uniquely qualified to provide Product Development support for all internal programs owing to the pharmaceutical industrial experience in drug discovery and development of its members and the state-of-the art facilities and equipment available to them. The following is an inventory of capabilities and assays that Core C can perform in-house with minimal outsourcing support.

Bioanalytical Chemistry

  • Staffed with experts in bioanalytical chemistry that have been trained in pharmaceutical industrial settings under the rigors of Good Laboratory Practice (GLP) guidelines.
  • Precise and accurate measurement of drug levels in various biological fluids (blood, plasma, urine, CSF, etc.) and a variety of animal tissues (brain, liver, lung, kidney, etc.).
  • Development and application of sensitive and selective bioanalytical methods utilizing HPLC coupled with triple-stage quadrupole mass spectrometry (LC/MS/MS).
  • Determination of drugs and metabolites in samples collected during in vivo animal PK/TK studies and in vitro drug metabolism (ADMET) investigations.

Drug Metabolism

Metabolic Stability in Liver Fractions

Evaluation of the metabolic stability of lead compounds in the presence (incubation) of liver fractions including microsomes, S9, and/or hepatocytes to estimate the in vivo half-lives in various species (mouse, rat, dog, monkey, and human).

Metabolite Identification and Characterization

Identification of metabolites of lead compounds in samples from in vitro experiments as well as samples collected during in vivo PK/TK studies • Characterization of major metabolites to assess their contributions to the toxicology and pharmacology of the lead compound.

Stability in Plasma

Incubation of compounds in plasma samples from various species (human, dog, rat, mouse, hamster, etc.) to assess their stability in plasma.

Plasma Protein Binding Studies

Assessment of plasma protein binding for all lead compounds using standard rapid equilibrium dialysis (RED) techniques with LC/MS/MS analysis in various species (mouse, rat, dog, monkey, and human).

CYP450 Inhibition Studies (Microsomes)

Determination of the CYP450 inhibition of test compounds by the five major isozymes (CYP1A2, 2C9, 2C19, 2D6, AND 3A4) of lead compounds using microsomes from various species (human, monkey, dog, rat, mice) with sample analysis by LC/MS/MS.

Drug Uptake by Cells, Cell Lines, Biological Fluids and Tissues

Drug Uptake and Anabolism by Cells/Cell Lines

Development of sensitive and selective methods for the quantitative determination of the intracellular levels of parent nucleoside/nucleotide compounds and the monophosphate (MP), diphosphate (DP), and triphosphate (TP) metabolites in samples from in vitro studies with various cells/cell lines.

Drug Uptake and Anabolism in Biological Fluids/Tissues

Quantitative determination of parent nucleoside/nucleotide compounds and their MP, DP, and TP metabolites in biological fluids (blood, plasma, urine, CSF, etc.) and various tissues (brain, liver, kidney, lung, etc.) from in vivo PK/TK studies.

In Vivo Pharmacokinetics/Toxicokinetics Studies

  • In vivo small animal PK/TK studies (mice, rats, etc.) using any of a number of modes of administration including oral, i.v., i.m., i.p., etc.) utilizing Yerkes Primate Research Center vivariums which are only a few steps away from the EIDD laboratories.
  • Estimation pharmacokinetic parameters (Cmax, Tmax, AUCs, t½) using standard pharmacokinetic software (Phoenix®WinNonlin® 6.3, Pharsight®) as well as compound distribution and concentration in specific target tissues ability to target specific tissues.

Chemistry, Manufacturing, and Control (CMC) Studies

Compound Solubility Determination

Routine determination of compound solubility at various pHs using Laser nephelometry (NEPHELOstar, BMG Lab Technologies).

Preliminary Dose Formulation Development

  • In-house development of routine formulations for use in dosage forms suitable for various dosing protocols (p.o., i.v., i.p., i.m., etc.) for in vivo PK/TK studies.
  • Development of dose formulations for IND-enabling studies through long term relationships with commercial formulators.

Analytical Chemistry

Support development programs with investigations of: compound stability (aqueous, organic, and buffers), structural confirmation/purity (HPLC/UV, LC/MS/MS), and compound formulation stability.