SpringerProtocols: Abstract: Absorption Screening Using the PAMPA Approach

Absorption Screening Using the PAMPA Approach

By: Jeffrey A. Ruell², Alex Avdeef²

Abstract

Full Text

Download PDF (624K)

The parallel artificial membrane permeability assay (PAMPA), as a passive-permeability screen, is an excellent alternative to cellular models for the earliest absorption, distribution, metabolism, and excretion (ADME) primary screening of research compounds. PAMPA’s popularity in the industry has risen rapidly. This chapter focuses on state-of-the-art PAMPA methods. Evidence will be cited demonstrating that as far as predicting passive permeability, PAMPA can outperform cellular models in speed, versatility, and especially cost, presenting a compelling and biologically relevant model of transport. The problem of low solubility of research compounds has been largely eliminated in the newest PAMPA variant, using a unique cosolvent method. This chapter also discusses how PAMPA has relevance in preformulation research. A detailed PAMPA protocol is presented, with step-by-step instructions for a popular version of the assay, using relatively inexpensive and readily obtainable components.

Affiliation(s): (2) pION INC, Woburn, MA

Book Title: Optimization in Drug Discovery: In Vitro Methods

Series: Methods in Pharmacology and Toxicology | Pub. Date: Aug-11-2004 | Page Range: 37-64 | DOI: 10.1385/1-59259-800-5:037

Subject: Pharmacology/Toxicology

Key Words: Oral absorption - permeability - artificial membranes - PAMPA - in vitro-in vivo correlations - unstirred water layer - cosolvent method for low-solubility assay

References

Download References

1.	Lipinski, C. A. (2002) Observation on current ADMET technology: no uniformity exists. Paper presented at the annual meeting of the Society of Biomolecular Screening, 24 September, The Hauge, The Netherlands.

2.	Di, L. and Kerns, E. H. (2003) Profiling drug-like properties in discovery research. Curr. Opin. Chem. Biol. 7, 402–408.

3.	Kerns, E. H. and Di, L. (2003) Pharmaceutical profiling in drug discovery. Drug Disc. Today 8, 316–323.

4.	Kansy, M., Senner, F., and Gubernator, K. (1998) Physicochemical high throughput screening: parallel artificial membrane permeability assay in the description of passive absorption processes. J. Med. Chem. 41, 1007–1010.

5.	Mueller, P., Rudin, D. O., Tien, H. T., and Westcott, W. C. (1962) Reconstitution of cell membrane structure in vitro and its transformation into an excitable system. Nature 194, 979–980.

6.	Tien, T. H. and Ottova, A. L. (2001) The lipid bilayer concept and its experimental realization: from soap bubbles, kitchen sink, to bilayer lipid membranes. J. Mem. Sci. 189, 83–117.

7.	Gutknecht, J. and Tosteson, D. C. (1973) Diffusion of weak acids across lipid membranes: effects of chemical reactions in the unstirred layers. Science 182, 1258–1261.

8.	Gutknecht, J., Bisson, M. A., and Tosteson, F. C. (1977) Diffusion of carbon dioxide through lipid bilayer membranes. Effects of carbonic anhydrase, bicarbonate, and unstirred layers. J. Gen. Physiol. 69, 779–794.

9.	Walter, A. and Gutknecht, J. (1984) Monocarboxylic acid permeation through lipid bilayer membranes. J. Mem. Biol. 77, 255–264.

10.	Antonenko, Y. N., Denisov, G. A., and Pohl, P. (1993) Weak acid transport across bilayer lipid membrane in the presence of buffers. Biophys. J. 64, 1701–1710.

11.	Xiang, T.-X. and Anderson, B. D. (1993) Diffusion of ionizable solutes across planar lipid bilayer membranes: boundary-layer pH gradients and the effect of buffers. Pharm. Res. 10, 1654–1661.

12.	Xiang, T.-X. and Anderson, B. D. (1994) Substituent contributions to the transport of substituted p-toluic acids across lipid bilayer membranes. J. Pharm. Sci. 83, 1511–1518.

13.	Mayer, P. T., Xiang, T.-X., and Anderson, B. D. (2000) Independence of substituent contributions to the transport of small-molecule permeants in lipid bilayers. AAPS PharmSci. 2, 1–13.

14.	Mayer, P. T. and Anderson, B. D. (2002) Transport across 1,9-decadiene precisely mimics the chemical selectivity of the barrier domain in egg lecithin bilayers. J. Pharm. Sci. 91, 640–646.

15.	Mayer, P. T., Xiang, T.-X., Niemi, R., and Anderson, B. D. (2003) A hydrophobicity scale for the lipid bilayer barrier domain from peptide permeabilities: nonadditivities in residue contributions. Biochemistry 42, 1624–1636.

16.	Mountz, J. M. and Tien, H. T. (1978) Photoeffects of pigmented lipid membranes in a micropourous filter. Photochem. Photobiol. 28, 395–400.

17.	Thompson, M., Lennox, R. B., and McClelland, R. A. (1982) Structure and electrochemical properties of microfiltration filter-lipid membrane systems. Anal. Chem. 54, 76–81.

18.	Cools, A. A. and Janssen, L. H. M. (1983) Influence of sodium ion-pair formation on transport kinetics of warfarin through octanol-impregnated membranes. J. Pharm. Pharmacol. 35, 689–691.

19.	Camenisch, G., Folkers, G., and van de Waterbeemd, H. (1997) Comparison of passive drug transport through Caco-2 cells and artificial membranes. Int. J. Pharm. 147, 61–70.

20.	Kansy, M., Fischer, H., Kratzat, K., Senner, F., Wagner, B., and Parrilla, I. (2001) High-throughput artificial membrane permeability studies in early lead discovery and development, in Pharmacokinetic Optimization in Drug Research, (Testa, B., van de Waterbeemd, H., Folkers, G., and Guy, R., eds.), Verlag Helvetica Chimica Acta, Zürich/Wiley-VCH, Weinheim, pp. 447–464.

21.	Avdeef, A. (2001) High-throughput measurements of solubility profiles, in in Pharmacokinetic Optimization in Drug Research, (Testa, B., van de Waterbeemd, H., Folkers, G., and Guy, R., eds.), Verlag Helvetica Chimica Acta, Zürich/Wiley-VCH, Weinheim, pp. 305–326.

22.	Faller, B. and Wohnsland, F. (2001) Physicochemical parameters as tools in drug discovery and lead optimization, in in Pharmacokinetic Optimization in Drug Research, (Testa, B., van de Waterbeemd, H., Folkers, G., and Guy, R., eds.), Verlag Helvetica Chimica Acta, Zürich/Wiley-VCH, Weinheim, pp. 257–274.

23.	Avdeef, A., Strafford, M., Block, E., Balogh, M. P., Chambliss, W., and Khan, I. (2001) Drug absorption in vitro model: filter-immobilized artificial membranes. 2. Studies of the permeability properties of lactones in piper methysticum forst. Eur. J. Pharm. Sci. 14, 271–280.

24.	Wohnsland, F. and Faller, B. (2001) High-throughput permeability pH profile and high-throughput alkane/water log P with artificial membranes. J. Med. Chem. 44, 923–930.

25.	Sugano, K., Hamada, H., Machida, M., Ushio, H., Saitoh, K., and Terada, K. (2001) Optimized conditions of bio-mimetic artificial membrane permeability assay. Int. J. Pharm. 228, 181–188.

26.	Sugano, K., Hamada, H., Machida, M., and Ushio, H. (2001) High throughput prediction of oral absorption: improvement of the composition of the lipid solution used in parallel artificial membrane permeability assay. J. Biomol. Screen. 6, 189–196.

27.	Sugano, K., Takata, N., Machida, M., Saitoh, K., and Terada, K. (2002) Prediction of passive intestinal absorption using bio-mimetic artificial membrane permeation assay and the paracellular pathway model. Int. J. Pharm. 241, 241–251.

28.	Sugano, K., Nabuchi, Y., Machida, M., and Aso, Y. (2003) Prediction of human intestinal permeability using artificial membrane permeability. Int. J. Pharm. 257, 245–251.

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

30.	Veber, D. F., Johnson, S. R., Cheng, H.-Y., Smith, B. R., Ward, K. W., and Kopple, K. D. (2002) Molecular properties that influence the oral bioavailability of drug candidates. J. Med. Chem. 45, 2615–2623.

31.	Di, L., Kerns, E. H., Fan, K., McConnell, O. J., and Carter, G. T. (2003) High throughput artificial membrane permeability assay for blood-brain barrier. Eur. J. Med. Chem. 38, 223–232.

32.	Avdeef, A. (2001) Physicochemical profiling (solubility, permeability, and charge state). Curr. Top. Med. Chem. 1, 277–351.

33.	Kerns, E. H. (2001) High throughput physicochemical profiling for drug discovery. J. Pharm. Sci. 90, 1838–1858.

34.	Kerns, E. H. and Di, L. (2002) Multivariate pharmaceutical profiling for drug discovery. Curr. Top. Med. Chem. 2, 87–98.

35.	Ruell, J. (2003) Membrane-based drug assays. Mod. Drug Disc. Jan., 28–30.

36.	Avdeef, A. (2003) High-throughput measurements of permeability profiles, in Drug Bioavailability: Estimation of Solubility, Permeability, Absorption and Bioavailability, (van de Waterbeemd, H., Lennernäs, H., and Artursson, P., eds.), Wiley-VCH, Weinheim, pp. 46–70.

37.	Faller, B. (2003) High-throughput physiochemical profiling: potential and limitations, in Analysis and Purification Methods in Combinatorial Chemistry. Wiley & Sons, New York, in press.

38.	Avdeef, A. (2003) Absorption and Drug Development. Wiley-Interscience, New York.

39.	Avdeef, A., Ruiz, A., Nalda, R., Ruell, J. A., Tsinman, O., Gonzalez, I., et al. (2004) PAMPA—a drug absorption in vitro model: 7. Comparing rat in situ, Caco-2, and PAMPA permeability of fluoroquinolones. Eur. J. Pharm. Sci., 21, 429–441.

40.	Nielsen, P. E. and Avdeef, A. (2004) PAMPA—a drug absorption in vitro model: 8. Apparent filter porosity and the unstirred water layer. Eur. J. Pharm. Sci. 21, in press.

41.	Ruell, J. A., Tsinman, K. L., and Avdeef, A. (2004) PAMPA—a drug absorption in vitro model: 5. Unstirred water layer in Iso-pH mapping assays and pK_a ^flux-optimized design (pOD-PAMPA). Eur. J. Pharm. Sci. 20, 393–402.

42.	Sugano, K., Nabuchi, Y., and Aso, Y. (2003) Transport mechanism of basic compounds through bio-mimetic artificial membrane. Poster presented at the Acad. Pharma. Sci. Technol. (APSTJ) Meeting, April, Kyoto, Japan.

43.	Lennernäs, H. (1998) Human intestinal permeability. J. Pharm. Sci. 87, 403–410.

44.	Partridge, W. M. (1991) Peptide Drug Delivery to the Brain. Raven Press, New York.

45.	Adson, A., Burton, P. S., Raub, T. J., Barsuhn, C. L., Audus, K. L., and Ho, N. F. H. (1995) Passive diffusion of weak organic electrolytes across Caco-2 cell monolayers: uncoupling the contributions of hydrodynamic, transcellular, and paracellular barriers. J. Pharm. Sci. 84, 1197–1204.

46.	Avdeef, A., Nielsen, P., and Tsinman, O. (2004) PAMPA—a drug absorption in vitro model: 11. Matching the in vivo unstirred water layer thickness by individual-well stirring in microtitre plates. Eur. J. Pharm. Sci. 21, in press.

47.	Pharma Algorithms, Toronto, Canada: http://www.ap-algorithms.com.

48.	Proulx, P. (1991) Structure-function relationships in intestinal brush border membranes. Biochim. Biophys. Acta 1071, 255–271.

49.	Ruell, J. A., Tsinman, O., and Avdeef, A. (2004) Acid-base cosolvent method for determining aqueous permeability of amiodarone, itra conazole, tamoxifen, ter fenadine and other very insoluble molecules. Chem. Pharm. Bull. 52, in press.

50.	Liu, H., Sabus, C., Carter, G. T., Avdeef, A., and Tischler, M. (2003) Solubilizer selection in the parallel artificial membrane permeability assay (PAMPA) for in vitro permeability measurement of low solubility compounds. Pharm. Res. 20, 1820–1826.

51.	Streng, W. H., Hsi, S. K., Helms, P. E., and Tan, H. G. H. (1984) General treatment of pH-solubility profiles of weak acids and bases and the effect of different acids on the solubility of a weak base. J. Pharm. Sci. 73, 1679–1684.

52.	Perrin, D. D. and Dempsey, B. (1974) Buffers for pH and Metal Ion Control. Chapman & Hall, London.

53.	Avdeef, A. and Bucher, J. J. (1978) Accurate measurements of the concentration of hydrogen ions with a glass electrode: calibrations using the Prideaux and other universal buffer solutions and a computer-controlled automatic titrator. Anal. Chem. 50, 2137–2142.

Comments (Loading...)

Post comment/View all comments