Gutbox technology

The stirring double-sink PAMPA method patented by pION Inc. (Billerica, MA) was employed to determine the permeability of compounds via PAMPA passive diffusion. The PAMPA lipid membrane consisted of an artificial membrane of a proprietary lipid mixture and dodecane (Pion Inc.), optimized to predict gastrointestinal tract passive diffusion permeability, and immobilized on a plastic matrix of a 96-well “donor” filter plate placed above a 96-well “acceptor” plate. A pH 7.4 solution was used in both donor and acceptor wells. The test articles, stocked in 10 mM DMSO solutions, were diluted to 0.05 mM in aqueous buffer (pH 7.4), and the concentration of DMSO was 0.5% in the final solution. During the 30 min permeation period at room temperature, the test samples in the donor compartment were stirred using the Gutbox technology (Pion Inc.) to reduce the unstirred water layer. The test article concentrations in the donor and acceptor compartments were measured using a UV plate reader (Nano Quant, Infinite 200 PRO, Tecan Inc., Männedorf, Switzerland). Permeabilitÿ calculations were performed using Pion Inc. software and were expressed in units of 10^–6 cm/s.

The stirring double-sink PAMPA method patented by pION Inc. (Billerica, MA) was employed to determine the permeability of compounds via PAMPA passive diffusion. The PAMPA lipid membrane consisted of an artificial membrane of a proprietary lipid mixture and dodecane (Pion Inc.), optimized to predict gastrointestinal tract (GIT) passive diffusion permeability, and was immobilized on a plastic matrix of a 96-well “donor” filter plate placed above a 96-well “acceptor” plate. A pH 7.4 solution was used in both donor and acceptor wells. The test articles, stocked in 10 mM DMSO solutions, were diluted to 0.05 mM in aqueous buffer (pH 7.4), and the concentration of DMSO was 0.5% in the final solution. During the 30 min permeation period at room temperature, the test samples in the donor compartment were stirred using the Gutbox technology (Pion Inc.) to reduce the unstirred water layer. The test article concentrations in the donor and acceptor compartments were measured using an UV plate reader (Nano Quant, Infinite 200 PRO, Tecan Inc., Mannedorf, Switzerland). Permeability calculations were performed using Pion Inc. software and were expressed in units of 10⁻⁶ cm s⁻¹. Compounds with values >100 are considered to have high permeability, compounds with values <100 but >10 are considered moderate permeability, and compounds with values <10 are considered to have low permeability.

The stirring double-sink PAMPA method patented by pION Inc. (Billerica, MA) was employed to determine the permeability of compounds via PAMPA passive diffusion. The PAMPA lipid membrane consisted of an artificial membrane of a proprietary lipid mixture and dodecane (Pion Inc.), optimized to predict gastrointestinal tract (GIT) passive diffusion permeability, was immobilized on a plastic matrix of a 96 well “donor” filter plate placed above a 96 well “acceptor” plate. A pH 7.4 solution was used in both donor and acceptor wells. The test articles, stocked in 10 mM DMSO solutions, were diluted to 0.05 mM in aqueous buffer (pH 7.4), and the concentration of DMSO was 0.5% in the final solution. During the 30-min permeation period at room temperature, the test samples in the donor compartment were stirred using the Gutbox technology (Pion Inc.) to reduce the unstirred water layer. The test article concentrations in the donor and acceptor compartments were measured using an UV plate reader (Nano Quant, Infinite 200 PRO, Tecan Inc., Männedorf, Switzerland). Permeability calculations were performed using Pion Inc. software and were expressed in units of 10^–6 cm/s.