Transport of thalidomide by the human intestinal caco-2 monolayers.
Studies in patients have indicated that the oral absorption of thalidomide is considerably variable at high doses (>200 mg/day). The aim of this study was to investigate the transport of racemic thalidomide using human colon cancer cell line (Caco-2) monolayers, which have been widely used to investigate drug permeability. A typical 21-day protocol was used to prepare Caco-2 monolayers. Thalidomide was determined by a validated high performance liquid chromatography method with ultraviolet detection. The integrity of Caco-2 monolayer was confirmed when the transepithelial electrical resistance (TEER) exceeded 300 Ohmz . cm2, and the leakage of 14C-manitol was <1% per hour. Uptake of thalidomide by Caco-2 cells was very limited (up to 2.1%). The transport of thalidomide appeared to be linear up to 1 hr. Our study indicated that the permeability coefficients (Papp) of thalidomide at 2.5-300 microM from the apical (AP) to basolateral (BL) and from BL to AP side was 2-6 x 10(-5) cm/sec, with a marked decrease in Papp values from AP to BL at increased thalidomide concentration. The transport of thalidomide was sodium-, temperature- and pH-dependent, as replacement of extracellular sodium chloride or reducing temperature and apical pH can result in significant decreases in the Papp values. Additional data indicated that transport of thalidomide is energy-dependent, as it was significantly (P < 0.05) inhibited by the ATP inhibitors, sodium azide and 2,4-dinitrophenol. In addition, DL-glutamic acid, cytidine, diprodomole, papaverine, quinidine, and cyclophosphamide significantly (P < 0.05) inhibited the transport of thalidomide, while the P-glycoprotein inhibitor verapamil and other nucleosides and nucleotides such as thymidine and guanine had no effect. These results indicated that thalidomide was rapidly transported by Caco-2 monolayers, and this might involve a saturable energy-dependent transporter.
Zhou
Shufeng
S
Department of Pharmacology and Clinical Pharmacology, Faculty of Medical and Health Sciences, the University of Auckland, Auckland, New Zealand.
Li
Yan
Y
Kestell
Phillip
P
Schafer
Peter
P
Chan
Eli
E
Paxton
James W
JW
D000255:Adenosine Triphosphate / Q000502:physiology; D001692:Biological Transport; D018938:Caco-2 Cells / Q000378:metabolism*; D004355:Drug Stability; D006801:Humans; D006863:Hydrogen-Ion Concentration; D013696:Temperature; D013792:Thalidomide / Q000493:pharmacokinetics*
D013792:Thalidomide; D000255:Adenosine Triphosphate