In Situ Artificial Membrane Permeation Assay under Hydrodynamic Control: Permeability-pH Profiles of Warfarin and Verapamil

Velický, Matěj; Bradley, Dan F.; Tam, Kin Yip; Dryfe, Robert A. W.

doi:10.1007/s11095-010-0150-6

Cited by 32 publications

(38 citation statements)

References 44 publications

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“…Warfarin, in contrast, diffuses freely in and out of the DPPC interface, in agreement with the behavior of deprotonated warfarin in atomistic simulations 44 . Lateral diffusion of both warfarin and brodifacoum occurs with a diffusivity on the order of 10 -6 cm 2 /s, comparing favorably with the membrane diffusion coefficient value of 1.611 ± 0.164 ×10 -6 cm 2 /s determined by artificial membrane permeation assay for warfarin 45 . We are not aware of any similar experimental measure for the much more rarely studied compound, brodifacoum, however, our diffusivity values for both species are comparable to experimental diffusion measurements for other small molecules 88, 89 .…”

Section: Structural Perturbation Of Dppc Bilayers Induced By Hydroxycsupporting

confidence: 67%

Structural perturbation of a dipalmitoylphosphatidylcholine (DPPC) bilayer by warfarin and its bolaamphiphilic analogue: A molecular dynamics study

Ayee

Roth

Akpa

2016

Journal of Colloid and Interface Science

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Compounds with nominally similar biological activity may exhibit differential toxicity due to differences in their interactions with cell membranes. Many pharmaceutical compounds are amphiphilic and can be taken up by phospholipid bilayers, interacting strongly with the lipid-aqueous interface whether or not subsequent permeation through the bilayer is possible. Bolaamphiphilic compounds, which possess two hydrophilic ends and a hydrophobic linker, can likewise undergo spontaneous uptake by bilayers. While membrane-spanning bolaamphiphiles can stabilize membranes, small molecules with this characteristic have the potential to create membrane defects via disruption of bilayer structure and dynamics. When compared to the amphiphilic therapeutic anticoagulant, warfarin, the bolaamphiphilic analogue, brodifacoum, exhibits heightened toxicity that goes beyond superior inhibition of the pharmacological target enzyme. We explore, herein, the consequences of anticoagulant accumulation in a dipalmitoylphosphatidylcholine (DPPC) bilayer. Coarse-grained molecular dynamics simulations reveal that permeation of phospholipid bilayers by brodifacoum causes a disruption of membrane barrier function that is driven by the bolaamphiphilic nature and size of this molecule. We find that brodifacoum partitioning into bilayers causes membrane thinning and permeabilization and promotes lipid flip-flop – phenomena that are suspected to play a role in triggering cell death. These phenomena are either absent or less pronounced in the case of the less toxic, amphiphilic compound, warfarin.

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Section: Structural Perturbation Of Dppc Bilayers Induced By Hydroxycsupporting

confidence: 67%

Structural perturbation of a dipalmitoylphosphatidylcholine (DPPC) bilayer by warfarin and its bolaamphiphilic analogue: A molecular dynamics study

Ayee

Roth

Akpa

2016

Journal of Colloid and Interface Science

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“…Permeation through the ABL is by passive diffusion. Hence, the ABL is a rate-limiting step for permeation of lipophilic compounds resulting in reduction of the apparent permeability (Hidalgo et al, 1991;Karlsson and Artursson, 1991;Ruell et al, 2003;Avdeef et al, 2004;Katneni et al, 2008;Velický et al, 2010), leading to reduced dynamic range and lower resolution in rank-ordering compound permeation. The ABL can also be a source of bias in determining the Michaelis-Menten transport kinetic K m because of the concentration gradient created within the ABL (Wilson and Dietschy, 1974;Balakrishnan et al, 2007) (Fig.…”

Section: Introductionmentioning

confidence: 99%

In vitro porcine blood–brain barrier model for permeability studies: pCEL-X software pKaFLUX method for aqueous boundary layer correction and detailed data analysis

Yusof

Avdeef²,

Abbott

2014

European Journal of Pharmaceutical Sciences

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In vitro blood-brain barrier (BBB) models from primary brain endothelial cells can closely resemble the in vivo BBB, offering valuable models to assay BBB functions and to screen potential central nervous system drugs. We have recently developed an in vitro BBB model using primary porcine brain endothelial cells. The model shows expression of tight junction proteins and high transendothelial electrical resistance, evidence for a restrictive paracellular pathway. Validation studies using small drug-like compounds demonstrated functional uptake and efflux transporters, showing the suitability of the model to assay drug permeability. However, one limitation of in vitro model permeability measurement is the presence of the aqueous boundary layer (ABL) resulting from inefficient stirring during the permeability assay. The ABL can be a rate-limiting step in permeation, particularly for lipophilic compounds, causing underestimation of the permeability. If the ABL effect is ignored, the permeability measured in vitro will not reflect the permeability in vivo. To address the issue, we explored the combination of in vitro permeability measurement using our porcine model with the pKa(FLUX) method in pCEL-X software to correct for the ABL effect and allow a detailed analysis of in vitro (transendothelial) permeability data, Papp. Published Papp using porcine models generated by our group and other groups are also analyzed. From the Papp, intrinsic transcellular permeability (P0) is derived by simultaneous refinement using a weighted nonlinear regression, taking into account permeability through the ABL, paracellular permeability and filter restrictions on permeation. The in vitro P0 derived for 22 compounds (35 measurements) showed good correlation with P0 derived from in situ brain perfusion data (r(2)=0.61). The analysis also gave evidence for carrier-mediated uptake of naloxone, propranolol and vinblastine. The combination of the in vitro porcine model and the software analysis provides a useful tool to better predict BBB permeability in vivo and gain better mechanistic information about BBB permeation.

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“…It has been shown that controlled hydrodynamics or stirring of the membrane improved the predictive power of in vitro permeability assays, thus mimicking the diffusion/convection conditions in a human intestine [69,70]. The traditional view of the widely accepted pH-partition hypothesis has also been challenged using a hydrodynamic approach to permeability [71]. It is interesting to note that several research groups independently pursued studies on the electrochemical response of SLM systems under hydrodynamic control.…”

Section: Voltammetry In Membrane Permeability Studiesmentioning

confidence: 99%

Use of voltammetry for in vitro equilibrium and transport studies of ionisable drugs

et al. 2014

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In this review, we will briefly outline the voltammetric investigations of the transfer of ionisable drugs at the interface between two immiscible electrolyte solutions. The voltammetric techniques enable the determination of some key in vitro properties of ionisable drugs, including partition coefficient, diffusioncoefficient and membrane permeability. Some successful applications will be highlighted, together with the background methodologies.

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In Situ Artificial Membrane Permeation Assay under Hydrodynamic Control: Permeability-pH Profiles of Warfarin and Verapamil

Cited by 32 publications

References 44 publications

Structural perturbation of a dipalmitoylphosphatidylcholine (DPPC) bilayer by warfarin and its bolaamphiphilic analogue: A molecular dynamics study

Structural perturbation of a dipalmitoylphosphatidylcholine (DPPC) bilayer by warfarin and its bolaamphiphilic analogue: A molecular dynamics study

In vitro porcine blood–brain barrier model for permeability studies: pCEL-X software pKaFLUX method for aqueous boundary layer correction and detailed data analysis

Use of voltammetry for in vitro equilibrium and transport studies of ionisable drugs

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