Caco-2 cells are used as an in-vitro model of the human intestinal epithelium and permit assessment of the intestinal permeability of potential drugs. In the bidirectional monolayer transport format, a confluent monolayer of polarized epithelial cells is established on a semi-permeable filter separating the two chambers of a Transwell apparatus. Test compound is added to either the apical or basolateral side of a confluent monolayer of Caco-2 cells and permeability is measured by monitoring the appearance of the test compound on the opposite side of the monolayer using appropriate analytical assays such as ELISA, HPLC etc.
Drug that is absorbed orally is transported via the portal circulation to the liver. In the liver the drug is oxidized by different isoforms of cytochrome P450 family. Drug metabolism is important for drug properties such as: drug stability and drug-drug interactions.
• CYP450 Inhibition (drug-drug interactions) – To assess the potential of the test compound to inhibit the main cytochrome P450 isoforms, CYP1A, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6 and CYP3A4. CYP inhibition will be evaluated using fluorescence substrate that will be metabolized by microsomes expressing a specific recombinant human P450 into highly fluorescent metabolite. We deliver IC50 for the test compound for each isoform.
• CYP450 Induction (drug- drug interaction) - identifies the potential of test compounds to induce CYP1A2, CYP2B6 or CYP3A4 in fresh cultured human hepatocytes by evaluating catalytic activity or protein and mRNA levels of P450 isoforms. We deliver fold of induction about vehicle control.
The solubility of a compound is an important factor in determining its absorption from the gastrointestinal tract and ultimately its oral bioavailability. Poor solubility can also limit the quality of the data generated in other in-vitro assays.
• We deliver absorbance spectra of compound in aqueous buffer or in formulation buffer.
Drug toxicity is a crucially important drug property. For each assay we deliver IC50 of tested compound. This assay can be performed on several cell lines including Caco2, HepG2, hepatocytes, and others.
• ATP consumption assays
• Cytotoxicity - WST1/MTT
• Biochemical Toxicology Assays- LDH activity, ALT activity, AST activity
Cardio toxicity (hERG)
The human ether-a-go-go related gene (hERG) encodes the voltage gated potassium channel in the heart (IKr) which is involved in cardiac re-polarization. As it might be sensitive to the tested compound its cardio toxic effect the undesired blockage of hERG must be determine. hERG inhibition will be determined using fluorescence polarization (FP), in which a red-shifted fluorescent tracer is displaced from the hERG channel by compounds that bind to the channel. When the tracer is bound to the channel the FP value is high, and when displaced by hERG binding compounds the FP value is low. We deliver IC50 for the test compound.