Several in vitro assays have been developed to evaluate the gastrointestinal absorption of compounds. Our aim was to compare 3 of these methods: 1) the bio-mimetic artificial membrane permeability assay (BAMPA) method, which offers a high-throughput, noncellular approach to the measurement of passive transport; 2) the traditional Caco-2 cell assay, the use of which as a high-throughput tool is limited by the long cell differentiation time (21 days); and 3) The BioCoat™ high-throughput screening Caco-2 Assay System, which reduces Caco-2 cell differentiation to 3 days. The transport of known compounds (such as cephalexin, propranolol, or chlorothiazide) was studied at pH 7.4 and 6.5 in BAMPA and both Caco-2 cell models. Permeability data obtained was correlated to known values of human absorption. Best correlations (r = 0.9) were obtained at pH 6.5 for BAMPA and at pH 7.4 for the Caco-2 cells grown for 21 days. The Caco-2 BioCoat™ HTS Caco-2 Assay System does not seem to be adequate for the prediction of absorption. The overall results indicate that BAMPA and the 21-day Caco-2 system can be complementary for an accurate prediction of human intestinal absorption.
1. Bohets H, Annaert P, Mannens G, Van Beijsterveldt L, Anciaux K, Verboven P, et al: Strategies for absorption screening in drug discovery and development. Curr Top Med Chem2001; 1: 367-383.
2.
2. Hidalgo IJ: Assessing the absorption of new pharmaceuticals. Curr Top Med Chem2001; 1: 385-401.
3.
3. Balimane PV, Chong S, Morrison RA: Current methodologies used for evaluation of intestinal permeability and absorption. J Pharmacol Toxicol Methods2000; 44: 301-312.
4.
4. Sugano K, Hamada H, Machida M, Ushio H: High throughput prediction of oral absorption: improvement of the composition of the lipid solution usedin parallel artificial membrane permeation assay. J Biomol Screen2001; 6: 189-196.
5.
5. Sugano K, Takata N, Machida M, Saitoh K, Terada K: Prediction of passive intestinal absorption using bio-mimetic artificial membrane permeation assay and the paracellular pathway model. Int J Pharm2002; 241: 241-251.
6.
6. Zhu C, Jiang L, Chen TM, Hwang KK: A comparative study of artificial membrane permeability assay for high throughput profiling of drug absorption potential. Eur J Med Chem2002; 37: 399-407.
7.
7. Sugano K, Hamada H, Machida M, Ushio H, Saitoh K, Terada K: Optimized conditions of bio-mimetic artificial membrane permeation assay. Int J Pharm2001; 228: 181-188.
8.
8. Ganapathy ME, Brandsch M, Prasad PD, Ganapathy V, Leibach FH: Differential recognition of beta-lactam antibiotics by intestinal and renal peptide transporters, PEPT 1 and PEPT 2. J Biol Chem1995; 270: 25672-25677.
9.
9. Chu XY, Sanchez-Castano GP, Higaki K, Oh DM, Hsu CP, Amidon GL: Correlation between epithelial cell permeability of cephalexin and expression of intestinal oligopeptide transporter. J Pharmacol Exp Ther2001; 299: 575-582.
10.
10. Walker D, Thwaites DT, Simmons NL, Gilbert HJ, Hirst BH: Substrate upregulation of the human small intestinal peptide transporter, hPepT1. J Physiol1998; 507: 697-706.
11.
11. Griffiths NM, Hirst BH, Simmons NL: Active intestinal secretion of the fluoroquinolone antibacterials ciprofloxacin, norfloxacin and pefloxacin: a common secretory pathway?J Pharmacol Exp Ther1994; 269: 496-502.
12.
12. Flanagan SD, Takahashi LH, Liu X, Benet LZ: Contributions of saturable active secretion, passive transcellular, and paracellular diffusion to the overall transport of furosemide across adenocarcinoma (Caco-2) cells. J Pharm Sci2002; 91: 1169-1177.
13.
13. Chiou WL, Chung SM, Wu TC, Ma C: A comprehensive account on the role of efflux transporters in the gastrointestinal absorption of 13 commonly used substrate drugs in humans. Int J Clin Pharmacol Ther2001; 39: 93-101.
15. Putnam WS, Ramanathan S, Pan L, Takahashi LH, Benet LZ: Functional characterization of monocarboxylic acid, large neutral amino acid, bile acid and peptide transporters, and P-glycoprotein in MDCK and Caco-2 cells. J Pharm Sci2002; 91: 2622-2635.
16.
16. Fogh J, Fogh JM, Orfeo T: One hundred and twenty-seven cultured human tumor cell lines producing tumors in nude mice. J Natl Cancer Inst1977; 59: 221-226.
17.
17. Howell S, Kenny AJ, Turner AJ: A survey of membrane peptidases in two human colonic cell lines, Caco-2 and HT-29. Biochem J1992; 284(Pt 2):595-601.
18.
18. Briske-Anderson MJ, Finley JW, Newman SM: The influence of culture time and passage number on the morphological and physiological development of Caco-2 cells. Proc Soc Exp Biol Med1997; 214: 248-257.
19.
19. Artursson P: Cell cultures as models for drug absorption across the intestinal mucosa. Crit Rev Ther Drug Carrier Syst1991; 8: 305-330.
20.
20. Chong S, Dando SA, Morrison RA: Evaluation of Biocoat intestinal epithelium differentiation environment (3-day cultured Caco-2 cells) as an absorption screening model with improved productivity. Pharm Res1997; 14: 1835-1837.
21.
21. Yamashita S, Konishi K, Yamazaki Y, Taki Y, Sakane T, Sezaki H, et al: New and better protocols for a short-term Caco-2 cell culture system. J Pharm Sci2002; 91: 669-679.
22.
22. Ingels F, Deferme S, Destexhe E, Oth M, Van den MG, Augustijns P: Simulated intestinal fluid as transport medium in the Caco-2 cell culture model. Int J Pharm2002; 232: 183-192.
23.
23. 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 Sci2000; 10: 195-204.
24.
24. Markowska M,Oberle R, Juzwin S, Hsu CP, Gryszkiewicz M, Streeter AJ: Optimizing Caco-2 cell monolayers to increase throughput in drug intestinal absorption analysis. J Pharmacol Toxicol Methods2001; 46: 51-55.
