Determination of effective concentrations of drug absorption enhancers using in vitro and ex vivo models

Abstract

Background: Achievement of an effective concentration of the pharmaceutically active ingredient in the blood and/or at the target site is an important aspect in the formulation of drugs and therefore needs to be quantified. Any concentration above therapeutic levels can cause toxic effects whereas low concentrations can be sub-therapeutic. This paper investigated different concentrations of selected commercially sourced analytical-grade pure chemicals as potential drug absorption enhancers in vitro and ex vivo to determine the lowest effective concentrations for optimizing drug absorption in oral dosage forms.

Methods: Recombinant cytochrome (CYP) 3A4 enzyme and recombinant p-glycoprotein membrane models were utilized for the investigation of in vitro inhibitory effects of drug absorption enhancers. Promega (2015) protocols were adopted for both assays. The everted porcine intestinal ex vivo model was employed for assessing effects of the drug absorption enhancers on the absorption of propranolol.

Results: The lowest effective CYP3A4 inhibitory concentrations were observed for curcumin (75µM and 100 µM), quercetin (75 and 100 µM) and glycyrrhizic acid (50 µM) while the most effective p-glycoprotein (P-gp) inhibition concentrations were curcumin (10, 15, 25, 50, 75 and 100 µM), sinomenine (50, 75, and 100 µM), quercetin (75 and 100 µM) and naringin (50 µM). Additive effects were observed between combinations of quercetin (75 µM) and curcumin (100 µM); quercetin (75 µM) and curcumin (75 µM); quercetin (75 µM) and curcumin (50 µM), and quercetin (75 µM) with curcumin (10 µM), which increased the basal ex vivo absorption of propranolol from 1.24 ± 0.03 µg/mL to 5.19 ± 0.12 µg/mL, 4.17 ± 0.05 µg/mL, 3.86 ± 0.10 µg/mL, and 4.07± 0.05 µg/mL respectively, after 2 hours.

Conclusion: Incorporation of the drug absorption enhancers (e.g., curcumin and quercetin), at specific concentrations, in dosage forms could improve the bioavailability of the BCS Class III and IV drugs that are substrates of CYP3A4 and p-glycoprotein.

Keywords: Bioavailability; Cytochrome CYP3A4; Drug absorption enhancers; Ex vivo; In vitro; Minimum effective concentration; P-glycoprotein.