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A head-to-head Caco-2 assay comparison of the mechanisms of action of the intestinal permeation enhancers: SNAC and sodium caprate (C10).
Eur J Pharm Biopharm. 2020 Jul; 152:95-107.EJ

Abstract

Salcaprozate sodium (SNAC) and sodium caprate (C10) are the two leading intestinal permeation enhancers (PEs) in oral peptide formulations in clinical trials. There is debate over their mechanism of action on intestinal epithelia. The aims were: (i) to compare their effects on the barrier function by measuring transepithelial electrical resistance (TEER), permeability of FITC-4000 (FD4) across Caco-2 monolayers, and on immunohistochemistry of tight junction (TJ)-associated proteins; and (ii) to compare cellular parameters using conventional end-point cytotoxicity assays and quantitative high content analysis (HCA) of multiple sub-lethal parameters in Caco-2 cells. C10 (8.5 mM) reversibly reduced TEER and increased FD4 permeability across monolayers, whereas SNAC had no effects on either parameter except at cytotoxic concentrations. C10 exposure induced reorganization of three TJ proteins, whereas SNAC only affected claudin-5 localization. High concentrations of C10 and SNAC were required to cause end-point toxicology changes in vitro. SNAC was less potent than C10 at inducing lysosomal and nuclear changes and plasma membrane perturbation. In parallel, HCA revealed that both agents displayed detergent-like features that reflect initial membrane fluidization followed by changes in intracellular parameters. In conclusion, FD4 permeability increases in monolayers in response to C10 were in the range of concentrations that altered end-point cytotoxicity and HCA parameters. For SNAC, while HCA parameters were also altered in a similar overall pattern as C10, they did not lead to increased paracellular flux. These assays show that both agents are primarily surfactants, but C10 has additional TJ-opening effects. While these in vitro assays illucidate their epithelial mechanism of action, clinical experience suggests that they over-estimate their toxicology in the dynamic intestinal environment.

Authors+Show Affiliations

UCD School of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland; UCD Conway Institute, University College Dublin, Belfield, Dublin 4, Ireland.UCD Conway Institute, University College Dublin, Belfield, Dublin 4, Ireland; UCD School of Chemistry, University College Dublin, Belfield, Dublin 4, Ireland.UCD School of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland.UCD Conway Institute, University College Dublin, Belfield, Dublin 4, Ireland; UCD School of Chemistry, University College Dublin, Belfield, Dublin 4, Ireland.School of Pharmacy, Institut Galien, CNRS, Univ. Paris-Sud, Univ. Paris-Saclay, 92290 Châtenay-Malabry, France.Drug Product Development, Small Molecules Oral Platform, Sanofi Research and Development, Montpellier, France.UCD School of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland; UCD Conway Institute, University College Dublin, Belfield, Dublin 4, Ireland. Electronic address: david.brayden@ucd.ie.

Pub Type(s)

Comparative Study
Journal Article

Language

eng

PubMed ID

32387703

Citation

Twarog, Caroline, et al. "A Head-to-head Caco-2 Assay Comparison of the Mechanisms of Action of the Intestinal Permeation Enhancers: SNAC and Sodium Caprate (C10)." European Journal of Pharmaceutics and Biopharmaceutics : Official Journal of Arbeitsgemeinschaft Fur Pharmazeutische Verfahrenstechnik E.V, vol. 152, 2020, pp. 95-107.
Twarog C, Liu K, O'Brien PJ, et al. A head-to-head Caco-2 assay comparison of the mechanisms of action of the intestinal permeation enhancers: SNAC and sodium caprate (C10). Eur J Pharm Biopharm. 2020;152:95-107.
Twarog, C., Liu, K., O'Brien, P. J., Dawson, K. A., Fattal, E., Illel, B., & Brayden, D. J. (2020). A head-to-head Caco-2 assay comparison of the mechanisms of action of the intestinal permeation enhancers: SNAC and sodium caprate (C10). European Journal of Pharmaceutics and Biopharmaceutics : Official Journal of Arbeitsgemeinschaft Fur Pharmazeutische Verfahrenstechnik E.V, 152, 95-107. https://doi.org/10.1016/j.ejpb.2020.04.023
Twarog C, et al. A Head-to-head Caco-2 Assay Comparison of the Mechanisms of Action of the Intestinal Permeation Enhancers: SNAC and Sodium Caprate (C10). Eur J Pharm Biopharm. 2020;152:95-107. PubMed PMID: 32387703.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A head-to-head Caco-2 assay comparison of the mechanisms of action of the intestinal permeation enhancers: SNAC and sodium caprate (C10). AU - Twarog,Caroline, AU - Liu,Kai, AU - O'Brien,Peter J, AU - Dawson,Kenneth A, AU - Fattal,Elias, AU - Illel,Brigitte, AU - Brayden,David J, Y1 - 2020/05/06/ PY - 2019/12/12/received PY - 2020/04/15/revised PY - 2020/04/28/accepted PY - 2020/5/11/pubmed PY - 2020/5/11/medline PY - 2020/5/11/entrez KW - Excipient cytotoxicity KW - Intestinal permeation enhancers KW - Oral peptide delivery KW - SNAC KW - Sodium caprate KW - Tight junctions SP - 95 EP - 107 JF - European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V JO - Eur J Pharm Biopharm VL - 152 N2 - Salcaprozate sodium (SNAC) and sodium caprate (C10) are the two leading intestinal permeation enhancers (PEs) in oral peptide formulations in clinical trials. There is debate over their mechanism of action on intestinal epithelia. The aims were: (i) to compare their effects on the barrier function by measuring transepithelial electrical resistance (TEER), permeability of FITC-4000 (FD4) across Caco-2 monolayers, and on immunohistochemistry of tight junction (TJ)-associated proteins; and (ii) to compare cellular parameters using conventional end-point cytotoxicity assays and quantitative high content analysis (HCA) of multiple sub-lethal parameters in Caco-2 cells. C10 (8.5 mM) reversibly reduced TEER and increased FD4 permeability across monolayers, whereas SNAC had no effects on either parameter except at cytotoxic concentrations. C10 exposure induced reorganization of three TJ proteins, whereas SNAC only affected claudin-5 localization. High concentrations of C10 and SNAC were required to cause end-point toxicology changes in vitro. SNAC was less potent than C10 at inducing lysosomal and nuclear changes and plasma membrane perturbation. In parallel, HCA revealed that both agents displayed detergent-like features that reflect initial membrane fluidization followed by changes in intracellular parameters. In conclusion, FD4 permeability increases in monolayers in response to C10 were in the range of concentrations that altered end-point cytotoxicity and HCA parameters. For SNAC, while HCA parameters were also altered in a similar overall pattern as C10, they did not lead to increased paracellular flux. These assays show that both agents are primarily surfactants, but C10 has additional TJ-opening effects. While these in vitro assays illucidate their epithelial mechanism of action, clinical experience suggests that they over-estimate their toxicology in the dynamic intestinal environment. SN - 1873-3441 UR - https://www.unboundmedicine.com/medline/citation/32387703/A_head_to_head_Caco_2_assay_comparison_of_the_mechanisms_of_action_of_the_intestinal_permeation_enhancers:_SNAC_and_sodium_caprate__C10__ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0939-6411(20)30118-1 DB - PRIME DP - Unbound Medicine ER -