Abstract

The natural product goldenseal is a clinical inhibitor of CYP3A activity, as evidenced by a 40-60% increase in midazolam AUC after coadministration with goldenseal. The predominant goldenseal alkaloids berberine and (-)-β-hydrastine were previously identified as time-dependent CYP3A inhibitors using human liver microsomes. Whether these alkaloids contribute to the clinical interaction, as well as the primary anatomical site (hepatic vs. intestinal) and mode of CYP3A inhibition (reversible vs. time-dependent), remain uncharacterized. The objective of this study was to mechanistically assess the pharmacokinetic goldenseal-midazolam interaction using an integrated in vitro-in vivo-in silico approach. Using human intestinal microsomes, (-)-β-hydrastine was a more potent time-dependent inhibitor of midazolam 1'-hydroxylation than berberine (K_I and k_inact: 8.48 µM and 0.041 min^-1, respectively, vs. >250 µM and ~0.06 min^-1, respectively). Both the AUC and C_max of midazolam increased by 40-60% after acute (single 3 g dose) and chronic (1 g thrice daily x 6 days) goldenseal administration to healthy adults. These increases, coupled with a modest/no increase ({less than or equal to}23%) in half-life, suggested that goldenseal primarily inhibited intestinal CYP3A. A physiologically based pharmacokinetic interaction model incorporating berberine and (-)-β-hydrastine successfully predicted the observed goldenseal-midazolam interaction (within 20%) after both chronic and acute goldenseal administration. Simulations implicated (-)-β-hydrastine as the major alkaloid precipitating the interaction, primarily via time-dependent inhibition of intestinal CYP3A, after chronic and acute goldenseal exposure. Results highlight the potential interplay between time-dependent and reversible inhibition of intestinal CYP3A as the mechanism underlying natural product-drug interactions, even after acute exposure to the precipitant.

Significance Statement Natural products can alter the pharmacokinetics of an object drug, potentially resulting in increased off-target effects or decreased efficacy of the drug. The objective of the current work was to evaluate fundamental mechanisms underlying the clinically observed goldenseal-midazolam interaction. Results support the use of an integrated approach involving established in vitro assays, clinical evaluation, and PBPK modeling to elucidate the complex interplay between multiple phytoconstituents and various pharmacokinetic processes driving a drug interaction.