High-throughput permeability screening

Use of Caco-2 Cells in Drug Discovery 9.3.1 High-Throughput Permeability Screening... 

LC/MS/MS techniques with selective and sensitive detection methods make it possible to quantitatively analyze samples from Caco-2 cell and PAMPA buffer matrices. A high-throughput permeability screen with robust LC/MS technology can quickly generate information about structure-permeability relationships that are extremely valuable in the lead optimization phase for the selection of pre-clinical candidates with favorable oral bioavailability properties. 

In addition to screening compounds, PAMPA was recently applied for high-throughput formulation screening. For early in vivo toxicity studies, it is often necessary to rapidly develop an effective formulation to increase exposure. Such formulations are usually a combination of several excipients. Simultaneous assessment of solubility and permeability is required since formulation excipients may reduce the apparent permeability by reducing the free fraction of a drug [68]. 

These automated assays can be used for high-throughput ADME screening in early drug discovery. The double-sink PAMPA permeability assay mimics in vivo conditions by the use of a chemical sink in the acceptor wells and pH gradient in the donor wells. The use of the pION gut-box integrated on the deck has shortened the PAMPA assay incubation time to 30 minutes. The permeability coefficient and rank order correlate well with data obtained using the in vitro Caco-2 assay and in vivo permeability properties measured in rat intestinal perfusions. 

Fujikawa M, Ano R, Nakao K, Shimizu R and Akamatsu M. Relationships between structure and high-throughput screening permeability of diverse drugs with artificial membranes application to prediction of Caco-2 cell permeability. Bioorg Med Chem 2005 13 4721-32. 

When screening for absorption by passive membrane permeability, artificial membranes have the advantage of offering a highly reproducible and high-throughput system. Artificial membranes have been compared to Caco-2 cells and for passive... 

Kansy, M., Senner, F., Gubemator, K. Physicochemical high throughput screening parallel artificial membrane permeability assay in the description of passive absorption processes. J. Med. Chem. 1998, 41, 1007-1010. 

Sugano, K. Hamada, H. Machida, M. Ushio, H., High throughput prediction of oral absorption Improvement of the composition of the lipid solution used in parallel artificial membrane permeability assay, J. Biomolec. Screen. 6, 189-196 (2001). 

Alsenz J, Haenel E (2003) Development of a 7-day, 96-well Caco-2 permeability assay with high-throughput of P-gp screening in drug discovery. Eur J Pharm Biopharm 58 99-105. 

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