Skip to main content

Drug Permeability Profiling Using the Novel Permeapad® 96-Well Plate

Pharmaceutical Research volume 37, Article number: 93 (2020) Cite this article

Abstract

Purpose

Here, first experiences with a prototype tool for high throughput (passive) permeability profiling, a 96-well plate comprising the Permeapad® membrane, are reported. The permeabilities of a set of drugs were determined and compared to published measures of oral absorption, such as human fraction absorbed (Fa) and in vitro permeability values obtained using other tools.

Methods

The tool consists of a 96-well bottom and screen plate with the artificial, phospholipid-based barrier (Permeapad®) mounted between the plates’ lower and upper compartments. The permeability of 14 model compounds including high- and low-absorption drugs, cationic, anionic, zwitterionic and neutral molecules, was determined by quantifying the compounds’ transport over time, deriving the steady-state flux from the linear part of the cumulative curves and calculating the apparent permeability (Papp). The membrane structure was investigated in a high-resolution digital light microscope.

Results

The Permeapad® 96-well plate was found suited to distinguish high and low absorption drugs and yielded a hyperbolic correlation to Fa. The Papp values obtained were congruent with those determined with in-house prepared Permeapad® in the Franz cell set-up. Furthermore, good to excellent correlations were seen with Caco-2 permeability (R2 = 0.70) and PAMPA permeability (R2 = 0.89). Microscopic investigation of the Permeapad® barrier revealed the formation of phospholipid vesicles and myelin figures in aqueous environment.

Conclusion

The Permeapad® 96-well plate permeation set-up is a promising new tool for rapid and reproducible passive permeability profiling.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

USD 39.95

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Abbreviations

Fa :

Fraction absorbed in humans

HPLC:

High-performance liquid chromatography

PAMPA:

Parallel artificial membrane permeability assay

Papp :

Apparent permeability

PBS:

Phosphate buffered saline

PVDF:

Polyvinylidene fluoride

PVPA:

Phospholipid vesicle-based permeation assay

TFA:

Trifluoroacetic acid

TPSA:

Total polar surface area

UHPLC-UV:

Ultra-high-performance liquid chromatography with ultraviolet detection

UWL:

Unstirred water layer

References

  1. Sugano K, Kansy M, Artursson P, Avdeef A, Bendels S, Di L, et al. Coexistence of passive and carrier-mediated processes in drug transport. Nat Rev Drug Discov. 2010;9:597–614.

    Article  CAS  Google Scholar 

  2. Berben P, Bauer-Brandl A, Brandl M, Faller B, Flaten GE, Jacobsen A-C, et al. Drug permeability profiling using cell-free permeation tools: overview and applications. Eur J Pharm Sci. 2018;119:219–33.

    Article  CAS  Google Scholar 

  3. Kansy M, Senner F, Gubernator K. Physicochemical high throughput screening: parallel artificial membrane permeation assay in the description of passive absorption processes. J Med Chem American Chemical Society. 1998;41:1007–10.

    CAS  Google Scholar 

  4. Teksin ZS, Seo PR, Polli JE. Comparison of drug Permeabilities and BCS classification: three lipid-component PAMPA system method versus Caco-2 monolayers. AAPS J. 2010;12:238–41.

    Article  CAS  Google Scholar 

  5. Avdeef A, Artursson P, Neuhoff S, Lazorova L, Gråsjö J, Tavelin S. Caco-2 permeability of weakly basic drugs predicted with the double-sink PAMPA pKaflux method. Eur J Pharm Sci. 2005;24:333–49.

    Article  CAS  Google Scholar 

  6. Flaten GE, Dhanikula AB, Luthman K, Brandl M. Drug permeability across a phospholipid vesicle based barrier: a novel approach for studying passive diffusion. Eur J Pharm Sci. 2006;27:80–90.

    Article  CAS  Google Scholar 

  7. di Cagno M, Bibi HA, Bauer-Brandl A. New biomimetic barrier Permeapad™ for efficient investigation of passive permeability of drugs. Eur J Pharm Sci. 2015;73:29–34.

    Article  Google Scholar 

  8. Lasic DD. Structure of amphiphilic aggregates. In: D.D. Lasic, editor. Liposomes from Phys to Appl. Elsevier Amsterdam; 1993. p. 43–62.

  9. Volkova T V, Terekhova I V, Silyukov OI, Proshin AN, Bauer-Brandl A, Perlovich GL. Towards the rational design of novel drugs based on solubility, partitioning/distribution, biomimetic permeability and biological activity exemplified by 1,2,4-thiadiazole derivatives. Medchemcomm. The Royal Society of Chemistry; 2017;8:162–175.

  10. Bibi HA, di Cagno M, Holm R, Bauer-Brandl A. Permeapad™ for investigation of passive drug permeability: the effect of surfactants, co-solvents and simulated intestinal fluids (FaSSIF and FeSSIF). Int J Pharm. 2015;493:192–7.

    Article  CAS  Google Scholar 

  11. Volkova TV, Domanina EN, Kumeev RS, Proshin AN, Terekhova IV. The effect of different polymers on the solubility, permeability and distribution of poor soluble 1,2,4-thiadiazole derivative. J Mol Liq. 2018;269:492–500.

    Article  CAS  Google Scholar 

  12. Bibi HA, Holm R, Bauer-Brandl A. Simultaneous lipolysis/permeation in vitro model, for the estimation of bioavailability of lipid based drug delivery systems. Eur J Pharm Biopharm. 2017;117:300–7.

    Article  CAS  Google Scholar 

  13. Fong SYK, Martins SM, Brandl M, Bauer-Brandl A. Solid phospholipid dispersions for Oral delivery of poorly soluble drugs: investigation into Celecoxib incorporation and solubility-in vitro permeability enhancement. J Pharm Sci. 2016;105:1113–23.

    Article  CAS  Google Scholar 

  14. Jacobsen A-C, Elvang PA, Bauer-Brandl A, Brandl M. A dynamic in vitro permeation study on solid mono- and diacyl-phospholipid dispersions of celecoxib. Eur J Pharm Sci. 2019;127:199–207.

    Article  CAS  Google Scholar 

  15. Wu IY, Bala S, Škalko-Basnet N, di Cagno MP. Interpreting non-linear drug diffusion data: Utilizing Korsmeyer-Peppas model to study drug release from liposomes. Eur J Pharm Sci 2019;138:105026.

  16. Agafonov M, Volkova T, Kumeev R, Chibunova E, Terekhova I. Impact of pluronic F127 on aqueous solubility and membrane permeability of antirheumatic compounds of different structure and polarity. J Mol Liq. 2019;274:770–7.

    Article  CAS  Google Scholar 

  17. Volkova T, Kumeev R, Kochkina N, Terekhova I. Impact of Pluronics of different structure on pharmacologically relevant properties of sulfasalazine and methotrexate. J Mol Liq. 2019;289:111076.

    Article  CAS  Google Scholar 

  18. Hermanson GT. Chapter 10 - fluorescent probes. In: Hermanson GTBT-BT (third E, editor. Boston: Academic Press; 2013. p. 395–463.

  19. Brandl M, Tardi C, Drechsler M, Bachmann D, Reszka R, Bauer KH, et al. Three-dimensional liposome networks: freeze fracture electron microscopical evaluation of their structure and in vitro analysis of release of hydrophilic markers. Adv Drug Deliv Rev. 1997;24:161–4.

    Article  Google Scholar 

  20. Brandl M. Vesicular phospholipid gels: a technology platform. J Liposome Res Taylor & Francis. 2007;17:15–26.

    Article  CAS  Google Scholar 

  21. Sugano K. 5.19 - Artificial Membrane Technologies to Assess Transfer and Permeation of Drugs in Drug Discovery. In: Taylor JB, Triggle DJBT-CMCII, editors. Oxford: Elsevier; 2007. p. 453–487.

  22. Flaten GE, Bunjes H, Luthman K, Brandl M. Drug permeability across a phospholipid vesicle-based barrier: 2. Characterization of barrier structure, storage stability and stability towards pH changes. Eur J Pharm Sci. 2006;28:336–43.

    Article  CAS  Google Scholar 

  23. Karlsson J, Artursson P. A method for the determination of cellular permeability coefficients and aqueous boundary layer thickness in monolayers of intestinal epithelial ( Caco-2) cells grown in permeable filter chambers. Int J Pharm. 1991;71:55–64.

    Article  CAS  Google Scholar 

  24. Nielsen PE, Avdeef A. PAMPA—a drug absorption in vitro model: 8. Apparent filter porosity and the unstirred water layer. Eur J Pharm Sci. 2004;22:33–41.

    Article  CAS  Google Scholar 

  25. Wolk O, Markovic M, Porat D, Fine-Shamir N, Zur M, Beig A, Dahan A. Segmental-dependent intestinal drug permeability: development and model validation of In silico predictions Guided by In Vivo permeability values. J Pharm Sci Elsevier; 2019;108:316–325.

    Article  CAS  Google Scholar 

  26. Bibi HA, Holm R, Bauer-Brandl A. Use of Permeapad® for prediction of buccal absorption: a comparison to in vitro, ex vivo and in vivo method. Eur J Pharm Sci. 2016;93:399–404.

    Article  CAS  Google Scholar 

  27. Teksin ZS, Hom K, Balakrishnan A, Polli JE. Ion pair-mediated transport of metoprolol across a three lipid-component PAMPA system. J Control Release. 2006;116:50–7.

    Article  CAS  Google Scholar 

  28. Zhu C, Jiang L, Chen T-M, Hwang K-K. A comparative study of artificial membrane permeability assay for high throughput profiling of drug absorption potential. Eur J Med Chem. 2002;37:399–407.

    Article  CAS  Google Scholar 

  29. Artursson P, Karlsson J. Correlation between oral drug absorption in humans and apparent drug permeability coefficients in human intestinal epithelial (Caco-2) cells. Biochem Biophys Res Commun. 1991;175:880–5.

    Article  CAS  Google Scholar 

  30. Yazdanian M, Glynn SL, Wright JL, Hawi A. Correlating partitioning and Caco-2 cell permeability of structurally diverse small molecular weight compounds. Pharm Res. 1998;15:1490–4.

    Article  CAS  Google Scholar 

  31. Yamashita S, Furubayashi T, Kataoka M, Sakane T, Sezaki H, Tokuda H. Optimized conditions for prediction of intestinal drug permeability using Caco-2 cells. Eur J Pharm Sci. 2000;10:195–204.

    Article  CAS  Google Scholar 

  32. Ghartey-Tagoe EB, Morgan JS, Neish AS, Prausnitz MR. Increased permeability of intestinal epithelial monolayers mediated by electroporation. J Control Release. 2005;103:177–90.

    Article  CAS  Google Scholar 

  33. Kogan A, Kesselman E, Danino D, Aserin A, Garti N. Viability and permeability across Caco-2 cells of CBZ solubilized in fully dilutable microemulsions. Colloids Surfaces B Biointerfaces. 2008;66:1–12.

    Article  CAS  Google Scholar 

  34. Morrison RA, Chong S, Marino AM, Wasserman MA, Timmins P, Moore VA, et al. Suitability of Enalapril as a probe of the dipeptide transporter system: InVitro and in vivo studies. Pharm Res. 1996;13:1078–82.

    Article  CAS  Google Scholar 

  35. Antonescu IE, Rasmussen KF, Neuhoff S, Fretté X, Karlgren M, Bergström CAS, et al. The permeation of Acamprosate is predominantly caused by Paracellular diffusion across Caco-2 cell monolayers: a Paracellular modeling approach. Mol Pharm. American Chemical Society. 2019;16:4636–50.

    Article  CAS  Google Scholar 

  36. Pade V, Stavchansky S. Link between drug absorption solubility and permeability measurements in Caco-2 cells. J Pharm Sci Elsevier. 1998;87:1604–7.

    Article  CAS  Google Scholar 

  37. Takenaka T, Harada N, Kuze J, Chiba M, Iwao T, Matsunaga T. Application of a human intestinal epithelial cell monolayer to the prediction of Oral drug absorption in humans as a superior alternative to the Caco-2 cell monolayer. J Pharm Sci. 2016;105:915–24.

    Article  CAS  Google Scholar 

  38. Yee S. In vitro permeability across Caco-2 cells (colonic) can predict in vivo (small intestinal) absorption in man—fact or myth. Pharm Res. 1997;14:763–6.

    Article  CAS  Google Scholar 

  39. Zur M, Gasparini M, Wolk O, Amidon GL, Dahan A. The low/high BCS permeability class boundary: physicochemical comparison of Metoprolol and labetalol. Mol Pharm American Chemical Society. 2014;11:1707–14.

    Article  CAS  Google Scholar 

  40. Vertzoni M, Augustijns P, Grimm M, Koziolek M, Lemmens G, Parrott N, et al. Impact of regional differences along the gastrointestinal tract of healthy adults on oral drug absorption: an UNGAP review. Eur J Pharm Sci. 2019;134:153–75.

    Article  CAS  Google Scholar 

  41. Lüpfert C, Reichel A. Development and application of physiologically based pharmacokinetic-modeling tools to support drug discovery. Chem Biodivers. John Wiley & Sons, Ltd; 2005;2:1462–86.

Download references

Disclaimer

The sponsor has not taken any influence on study design, data interpretation or writing of the current manuscript.

Author information

Authors and Affiliations

  1. Drug Transport & Delivery Group, Department of Physics, Chemistry & Pharmacy, University of Southern Denmark, 5230, Odense, Denmark

    Ann-Christin Jacobsen, Sune Nielsen, Martin Brandl & Annette Bauer-Brandl

Authors
  1. Ann-Christin Jacobsen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sune Nielsen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Martin Brandl
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Annette Bauer-Brandl
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Annette Bauer-Brandl.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

ESM 1

(DOCX 54 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Jacobsen, AC., Nielsen, S., Brandl, M. et al. Drug Permeability Profiling Using the Novel Permeapad® 96-Well Plate. Pharm Res 37, 93 (2020). https://doi.org/10.1007/s11095-020-02807-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11095-020-02807-x

Key words

  • 96-well plate
  • artificial barrier
  • high throughput
  • intestinal absorption
  • microplate
  • permeability