The Lecithin and anionic lipids as an imitation of the lipid membrane in Parallel Artificial Membrane Permeation Assay (PAMPA) blood –brain barrier Models

Main Article Content

Svetlana Otasevic
Tanja Vojinovic

Keywords

Pampa; Blood-brain barrier; Lecithin; Anionic Lipids

Abstract

Background/Aim: One of the biggest challenges today is producing pharmaceutical forms of drugs for a successful treatment of pathological disorders of the central nervous system (CNS), bearing in mind that the brain tissue is highly selective when it comes to permeation of all substances, particularly drugs. In vitro Parallel Artificial Membrane Permeation Assay (PAMPA) drug permeability assay is an extremely important experimental model in the process of drug development. The aim of this study is to modify this model in order to test drug permeability through the blood-brain barrier. Methods: The substances with pharmacological activity tested in this study are the following: theophylline, sulfasalzine, risperidone, haloperidol, lidocaine and propranolol. We studied some of the key parameters which are important for good prediction of permeability – lipid composition and buffer composition. We also investigated the effect of co-solvents in the donor compartment and surfactants in the acceptor compartment, as well as the influence of the anionic lipid composition on drug permeability through the blood-brain barrier. As co-solvents, we used polyethylene glycol 200 (PEG 200) at concentrations of 1% and 0.24%, and as surfacants we applied sodim lauryl sulfate (SLS) at concentrations of 2% and 0.5%. In addition, the influence of lecithin and the anionic lipid composition was investigated and the following lipids were used for this purpose: 1,2-dioleoyl-sn-glycero-3-[phospho-L-serine] (PS18: 1), 1,2-dioleoyl-sn-glycero-3-phosphocholine (PC18: 1), and cholesterol. Results: The use of PEG 200 at concentrations of 1% and 0.24% proved to be good and allowed for a proper classification of the substances into those of high and low permeability. SLS at concentrations of 2 and 0.5% impaired the integrity of the lipid membrane, with the solution penetrating from the acceptor into the donor compartment. A brief experiment with methanol and SLS combined, showed that the membrane integrity remains preserved. The anionic lipid composition also proved favourable and the substances tested were properly classified into those of high and low permeability. Conclusion: The lipids that were used as an imitation of lipid membrane such as a 10% lecithin and combination of 1,2-dioleoyl-sn-glycero-3-[phospho-L-serine] (PS18: 1), 1,2-dioleoyl-sn-glycero-3-phosphocholine (PC18: 1) and cholesterol in the anionic-pampa model proved to be good and allowed for a proper classification of the substances into those of high and low permeability.

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References

[1] Kansy M, Senner F, Gubernator K.Physicochemical High Throughput Screening: Parallel Artificial Membrane Permeation Assay in the Description of Passive Absorption Processes, Journal Of Medicinal Chemistry. 41 (1998) 1007-1010.
[2] Pokrajac M, Farmakokinetika, Birograf Beograd (2007)
[3] Sugano K, Hamada H, Machida M, Ushio H, Saitoh K, Terada K. Optimized conditions of bio-mimetic artificial membrane permeation assay, International Journal Of Pharmaceutics. 228 (2001) 181-188.
[4] Di L, Kerns E, Fan K, McConnell O, Carter G. High throughput artificial membrane permeability assay for blood–brain barrier, European Journal Of Medicinal Chemistry. 38 (2003) 223-232.
[5] Berben P, Bauer-Brandl A, Brandl M, et al. Drug permeability profiling using cell-free permeation tools: Overview and applications, European Journal Of Pharmaceutical Sciences. 119 (2018) 219-233.
[6] Matsson P, Fenu L, Lundquist P, Wiśniewski J, Kansy M, Artursson, P. Quantifying the impact of transporters on cellular drug permeability. Trends in Pharmacological Sciences, 36 (5), (2015) pp.255-262.
[7] Ölander M, Wiśniewski JR, Matsson P, Lundquist P, Artursson P. The proteomeof filter- grown Caco-2 cells with a focus on proteins involved in drug disposition. J. Pharm. Sci. 105, (2016) 817–827.
[8] Sun D, Lennernas H, Welage S, at al. Comparison of human duodenum and Caco-2 gene expression profiles for 12,000 gene sequences tags and correlation with permeability of 26 drugs. Pharm. Res. 19, (2002) 1400–1416.
[9] Lundquist S, Renftlel M, Brillault J, Fenart L, Cecchelli R, Dehouck M. Prediction of drug transport through the blood–brain barrier in vivo: a comparison between two in vitro cell models, Pharm. Res. 19 (2002) 976–981.
[10] Lundquist S, Renftel M. The use of in vitro cell culture models for mechanistic studies and as permeability screens for the blood–brain barrier in the pharmaceutical industry – background and current status in the drug discovery process, Vascul. Pharmacol. 38 (2002) 355–364.
[11] Bicker J, Alves G, Fortuna A, Falcão A, Blood–brain barrier models and their relevance for a successful development of CNS drug delivery systems: A review, European Journal Of Pharmaceutics And Biopharmaceutics. 87 (2014) 409-432.
[12] Mensch J, Oyarzabal J, Mackie C, Augustijns P. In vivo, in vitro and in silico methods for small molecule transfer across the BBB, J. Pharm. Sci. 98 (2009) 4429–4468.
[13] M. Kansy, F. Senner, K. Gubernator. Physicochemical High Throughput Screening: Parallel Artificial Membrane Permeation Assay in the Description of Passive Absorption Processes, Journal Of Medicinal Chemistry. 41 (1998) 1007-1010.
[14] Teksin Z, Seo P, Polli J. Comparison of Drug Permeabilities and BCS Classification: Three Lipid-Component PAMPA System Method versus Caco-2 Monolayers, The AAPS Journal. 12 (2010) 238-241.
[15] Avdeef A. Absorption and drug development, John Wiley & Sons, Hoboken, N.J., 2012.
[16] Tsinman O, Tsinman K, Sun N, Avdeef A. Physicochemical Selectivity of the BBB Microenvironment Governing Passive Diffusion—Matching with a Porcine Brain Lipid Extract Artificial Membrane Permeability Model, Pharmaceutical Research. 28 (2010) 337-363.
[17] Chen X, Murawski A, Patel K, Crespi C, Balimane P, 2008. A Novel Design of Artificial Membrane for Improving the PAMPA Model. Pharmaceutical Research, Vol. 25, No. 7, July 2008