Passive Permeability Test Caco-2 Assays

These early investigations of compound pooling in the Caco-2 assay (Bu 2000 Laitinen 2003) used conventional gradient LC-MS/MS analytical methods with diversion of the first one to two minutes of the eluent to waste to avoid 

lb Drug-Drug Interaction Studies (CYP450 Assays) 

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The first step in this direction is to correlate the permeability values obtained in the Caco-2 cell lines (apical-basolateral direction, Pay) with the fraction of dose absorbed in vivo in rat For this proposal eight fluoroquinolones were assayed and the results found with Caco-2 cell lines were compared with those obtained in vivo in rat In the Caco-2 cell lines the permeability of the quinolones was evaluated at different initial concentration, in order to test for no linearities in the absorption process. For some of them, it was observed that a secretion system worked in the opposite direction to passive diffusion for this reason the permeability value used for the correlations, in the case of secretion, was the one obtained at the highest concentration of the quinolone, which corresponds to saturation of the secretion process. 

The parallel artificial membrane permeability assay (PAMPA) is a recent development in the area of artificial membranes that appears to offer considerable potential. Measuring the flux values (membrane permeation levels) of a range of test compounds by PAMPA and relating these values to the flux curves obtained in Caco-2 studies have shown good correlations, indicating that the PAMPA assay could be a good alternative to Caco-2 cells for the measurement of passively diffusing compounds. 

Artificial Systems (PAMPA) Some artificial systems are widely used as an alternative to Caco-2 cell culture to evaluate drug permeability. An example of these is the parallel artificial membrane permeation assay (PAMPA). PAMPA is usually performed in a 96-well microtiter plate format that consists of test wells fixed with an artificial hydrophobic membrane and reference wells (Kansy et al., 1998). Test compounds are applied at one side of the membrane, and the permeability rates are monitored at the receiving side. The properties of the membrane (lipophilicity and thickness) control the rate of permeability. Different membranes could be designed to mimic the passive... 

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