Custom Primary Cell Isolation
Primary hepatocytes are widely used in evaluating metabolism, drug-drug interactions, drug transporter activity, and potential toxicity of drug candidates. As a CompleteHepatocyteSolutionTM company, our goal is to provide hepatocytes of the highest quality to ensure our customers can produce reliable, consistent data. With IVAL, you aren’t restricted by a pre-set isolation calendar—let us know when you need a fresh hepatocyte preparation and we’ll prepare it to fit your schedule.
Metabolic Stability is a key drug property. It is important for drug administration regimen design as well as toxicity. Species comparison in metabolic stability allows the determination of which animal species is the most appropriate model to estimate human metabolic stability. The use of human primary hepatocytes allows the evaluation of metabolic stability as a result of both phase I oxidation and phase II conjugation.
Metabolic Pathway Identification (Reaction Phenotyping)
Predicting the potential for drug-drug interactions with co-administered compounds starts by understanding the metabolic pathway by which a compound is eliminated from the body. Compounds with a single route of elimination are at high risk of causing adverse events. To obviate any dangerous polypharmacy situations in patients, identifying the metabolic pathway(s) of a drug candidate is required by regulatory agencies prior to IND submission.
These studies are performed using liver microsomes in the presence of selective inhibitors for the major CYP isoforms, thus identifying the metabolic routes involved. Quantification of the parent compound and its metabolites are performed using HPLC-LC/MS, and the percent contribution of a specific CYP isoform towards metabolism of the test article is reported.
Metabolism Pathway Inhibition
A major mechanism of drug-drug interaction is the inhibition of drug metabolizing enzymes by a co-administered drug, thereby inhibiting the metabolism of co-administered drugs, which are substrates of the inhibited pathways. This can be performed using liver microsomes (human/animal) and hepatocytes readily available at IVAL. Choose between luminescent (CYP3A4 only) or LC/MS validated protocols. Analyses are performed in triplicate across seven concentrations of the test article. IC50 is reported.
Metabolism Pathway Induction
Enzyme induction is a major mechanism of pharmacokinetic drug-drug interactions, and has the potential to increase the metabolic clearance of a drug, thus compromising efficacy. Other concerns of induction are the increased production of active metabolites, tumor formation and toxicological consequences. The potential for adverse events due to enzyme induction is recognized by regulatory agencies worldwide and investigations into a compound’s potential to create or be subject to such an effect is considered an important part of the drug development regimen.
IVAL routinely performs hepatocyte-based induction studies in vitro, offering a large selection of human and animal hepatocytes as a test system, a choice of validated luminescent and/or LC/MS assays, and quick turnaround time.
In vitro testing using primary cells can quickly screen for new chemical entities with serious toxicology consequences. Using primary cells derived from human tissues, the relative risk can be determined in multiple organs over a population of donors.
The following in vitro cytotoxicity assays using the specified endpoints have been developed in our laboratory. With our ready inventory of human and cynomolgus monkey cells scheduling is not an issue. Contact us to customize your study today.
- Cell viability (luminescence based ATP assay)
- Cell growth and survival (MTT/WST-1 metabolism)
- Apoptosis (luminescence based caspase activity assay)
- Oxidative Stress (luminescence based glutathione assay)
Non-Hepatic Organ/IdMOCTM Technology
In typical in vitro systems, each cell type was studied in isolation, ignoring critical interactions between organs when metabolizing drugs. This led to the possibility of many factors such as inter-organ toxicity going unrecognized in drug evaluations.
The IdMOCTM technology, developed by IVAL, models in vivo multiple-organ interaction, thus allowing the evaluation of organ-specific effects of a drug and its metabolites. You may customize cell types as you need. Inquire us for further details.
Primary Human Cells
- Kidney proximal tubule cells
- Liver hepatocytes
- Small airway epithelial cells
- Neuronal cells
Pravastatin Uptake Assessment: 96 well cultures at a cell density of 0.5 million hepatocytes/mL (50,000 hepatocytes/well) were used in the Pravastatin Uptake Assessment. After approximately 20-24 hours in culture, the hepatocytes were treated with 1 µM of Pravastatin with and without Rifampin for a time duration of 0,1, 2, 3, 4, and 5 minutes. Values reflect the mean of triplicate treatments (N=3). The metabolites were identified and analyzed using LCMS/MS.