Metabolite Profiling & Identification in vitro / in vivo
Aim: To identify in vitro and/or in vivo the metabolites of test compound, and to compare across species the in vitro-in vivo correlation and metabolic profiles.
Background information: Metabolite profiling of compounds at early stages in the Drug Discovery process is relevant to detect metabolites which are unique to humans compared with animal species, or metabolites present at disproportionately higher levels in humans than in animal species. In these cases, it may be necessary to assess the preclinical safety of the metabolite detected.
On the other hand, the identification of the metabolic liability of the test compound is important in directing chemistry. If a compound is cleared very rapidly, then identifying which functional groups are undergoing metabolism will be valuable in understanding how the chemical structure may be modifed to reduce compound biotransformation.
Turnaround time: depends on depth level of data and interpretation required
Matrices: (flexible options depending on the needs)
- In vitro metabolism: liver microsomes, hepatocytes, S9, recombinant enzymes or extrahepatic tissues
- In vivo metabolism: urine, plasma or tissue samples
- Proposed chemical structures for metabolites found based on their theoretical pseudomolecular mass m/z and MS/MS fragmentation pattern
- Detection of the key positions in a molecule that undergo biotransformation
- Proportion of metabolites observed assuming similar MS response for parent compound and metabolites if standards are not available.
- Comparison of metabolic profile among species to detect potential differences
- Structural confirmation of proposed metabolites if standards are available