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Published on: April 9, 2014
Rapid assessment of P-glycoprotein inhibition and induction in vitro
Michael D Perloff1, Elke Störmer, Lisa L von Moltke
1Department of Pharmacology and Experimental Therapeutics, Tufts University School of Medicine, 136 Harrison Avenue, Boston, Massachusetts 02111, USA.
Purpose:
Using rhodamine123 (RH123) cell exclusion. 17 clinically used compounds were screened for their inhibitory effect on P-glycoprotein (P-gp), which was compared with the drugs' inhibitory activity against CYP3A4. The same assay was used to study induction of P-gp activity.
Methods:
P-gp inhibition was assessed using RH123 accumulation into LS180V cells as well as Rh123 transport across Caco-2 mono-layers. Inhibition of CYP3A4 was determined in human liver microsomes using triazolam-4-hydroxylation. Induction of P-gp expression and activity was measured using western blot analysis and RH123 accumulation into LS180V cells, respectively.
Results:
The observed inhibition of RH123 cell exclusion ranged from little or no effect (digoxin, indinavir, fexofenadine) up to a nearly 10-fold increase in RH 123 accumulation (ivermectin, terfenadine). No correlation between P-gp and CYP3A4 inhibition was observed. The rank order in P-gp inhibitory potency for terfenadine, verapamil, ritonavir. and indomethacin was identical in both LS180V and Caco-2 models. Ritonavir and St. John's wort extract showed a concentration-dependent P-gp induction, with good correlation between western blot analysis and RH123 accumulation.
Conclusions:
The RH123 accumulation assay in LS180V cells can be used as a valuable screening tool to study both inhibition and induction of P-gp activity and expression. This assay has the potential to predict P-gp-mediated alterations in intestinal absorption of drugs.
