In Vitro Metabolism & Identification
Understanding how an investigational compound metabolizes is an important part of developing a strong regulatory package and strategy for administration of the compound. Frontage’s in vitro ADME team provides supportive studies to perform in vitro metabolism and cell culture with dedicated analytical support (UFLC-MS/MS, Q-Trap/MS/MS, HPLC) and software tools.
- Reaction phenotyping (enzymology) is an assessment of the enzymes responsible for metabolizing a potential new drug in human liver providing integral information.
- CYP450 inhibition: Inhibition of cytochrome P450 enzymes is a significant cause of metabolically-based drug-drug interactions in vivo. Therefore, an assessment of the CYP450 inhibition potential is essential for compound development and clinical research.
- CYP450 induction: As compounds are administered over longer periods of time, some can induce increased levels of metabolic enzymes. This can lead to decreased efficacy and possible formation of toxic metabolites. Frontage performs enzyme induction studies using cultured human hepatocytes, which is in line with FDA recommended practices. After exposure, the effect of the test compound can be assessed using RT-PCR.
- P-gp interactions: Transporter-based drug-drug interactions in vivo are being increasingly reported in the scientific literature. These interactions can affect permeability and distribution of compounds. Frontage can assess interactions with P-gp (as both substrate and inhibitor) using Caco-2 human adenocarcinoma cell lines, as recommended by the FDA.
Other in vitro drug development studies conducted at Frontage include:
- Species comparison of metabolism provides essential metabolic information early in the development process. The rate and metabolic route(s) of metabolism for a new compound can be readily compared between species and compared to human results to aid in the selection of appropriate animal models for safety assessments of the compound in development. In particular, we are experts at isolating hepatocytes and using the freshly isolated hepatocyte model for studies of this type, which is the preferred model.
- Plasma protein binding studies are required to aide in the interpretation of in vivo pharmacokinetic and toxicokinetic information. Frontage can perform studies to assess plasma protein binding across a range of species using either equilibrium dialysis, ultrafiltration or ultracentrifugation.