Version 1
: Received: 29 May 2024 / Approved: 30 May 2024 / Online: 30 May 2024 (13:45:41 CEST)
How to cite:
Pires, C. L.; Rosa, S.; Batista de Carvalho, A. L. M.; Ferreira, L.; Marques, M. P. M.; Moreno, M. J. Re-Use of Caco-2 Monolayers for a Higher Throughput Assessment of Drug Permeability – Methodology and Validation for Passive Permeation. Preprints2024, 2024052057. https://doi.org/10.20944/preprints202405.2057.v1
Pires, C. L.; Rosa, S.; Batista de Carvalho, A. L. M.; Ferreira, L.; Marques, M. P. M.; Moreno, M. J. Re-Use of Caco-2 Monolayers for a Higher Throughput Assessment of Drug Permeability – Methodology and Validation for Passive Permeation. Preprints 2024, 2024052057. https://doi.org/10.20944/preprints202405.2057.v1
Pires, C. L.; Rosa, S.; Batista de Carvalho, A. L. M.; Ferreira, L.; Marques, M. P. M.; Moreno, M. J. Re-Use of Caco-2 Monolayers for a Higher Throughput Assessment of Drug Permeability – Methodology and Validation for Passive Permeation. Preprints2024, 2024052057. https://doi.org/10.20944/preprints202405.2057.v1
APA Style
Pires, C. L., Rosa, S., Batista de Carvalho, A. L. M., Ferreira, L., Marques, M. P. M., & Moreno, M. J. (2024). Re-Use of Caco-2 Monolayers for a Higher Throughput Assessment of Drug Permeability – Methodology and Validation for Passive Permeation. Preprints. https://doi.org/10.20944/preprints202405.2057.v1
Chicago/Turabian Style
Pires, C. L., Maria Paula M. Marques and Maria João Moreno. 2024 "Re-Use of Caco-2 Monolayers for a Higher Throughput Assessment of Drug Permeability – Methodology and Validation for Passive Permeation" Preprints. https://doi.org/10.20944/preprints202405.2057.v1
Abstract
Evaluation of drug permeability using cell monolayers is an important in vitro assay in the process of drug development, to identify compounds with favourable pharmacokinetics that may proceed to in vivo studies. When allowed to differentiate on permeable filters for at least 21 days, monolayers of Caco-2 cells develop morphological and functional properties similar to those of the intestinal epithelium. Permeability through Caco-2 monolayers is thus particularly relevant to assess drug absorption after oral administration. The major drawback of this assay is its very low throughput, with the cell monolayers being used in a single permeability assay. We have previously proposed a protocol to allow the re-use of the Caco-2 monolayer on additional permeability assays during the stable period and shown that cell monolayer integrity is maintained. In this work we evaluate the effect of re-use on the permeability of several reference compounds that permeate through passive pathways and show that the cell monolayers can be re-used twice in additional permeability assays. Preliminary results were also obtained regarding active transport pathways, and a new methodology is proposed to quantify the contribution from passive and active transport. This is a major step towards the full validation of the re-use methodology proposed, which at least triplicates the throughput of this important in vitro assay.
Keywords
Permeability assay; pharmacokinetics; Caco-2 monolayers; re-use; high throughput; passive diffusion; active transport
Subject
Medicine and Pharmacology, Pharmacology and Toxicology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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