Effect of P. corylifolia on the pharmacokinetic profile of tofacitinib and the underlying mechanism

Abstract

This work aimed to explore the mechanisms underlying the interaction of the active furanocoumarins in P. corylifolia on tofacitinib both in vivo and in vitro. The concentration of tofacitinib and its metabolite M8 was determined using UPLC-MS/MS. The peak area ratio of M8 to tofacitinib was calculated to compare the inhibitory ability of furanocoumarin contained in the traditional Chinese medicine P. corylifolia in rat liver microsomes (RLMs), human liver microsomes (HLMs) and recombinant human CYP3A4 (rCYP3A4). We found that bergapten and isopsoralen exhibited more significant inhibitory activity in RLMs than other furanocoumarins. Bergapten and isopsoralen were selected to investigate tofacitinib drug interactions in vitro and in vivo. Thirty rats were randomly allocated into 5 groups (n = 6): control (0.5% CMC-Na), low-dose bergapten (20 mg/kg), high-dose bergapten (50 mg/kg), low-dose isopsoralen (20 mg/kg) and ketoconazole. 10 mg/kg of tofacitinib was orally intervented to each rat and the concentration level of tofacitinib in the rats were determined by UPLC-MS/MS. More imporrantly, the results showed that bergapten and isopsoralen significantly inhibited the metabolism of tofacitinib metabolism. The AUC_(0-t), AUC_(0-∞), MRT_(0-t), MRT_(0-∞) and Cmax of tofacitinib increased in varying degrees compared with the control group (all p < 0.05), but CLz/F decreased in varying degrees (p < 0.05) in the different dose bergapten group and isopsoralen group. Bergapten, isopsoralen and tofacitinib exhibit similar binding capacities with CYP3A4 by AutoDock 4.2 software, confirming that they compete for tofacitinib metabolism. P. corylifolia may considerably impact the metabolism of tofacitinib, which can provide essential information for the accurate therapeutic application of tofacitinib.

Keywords: P. corylifolia; cytochrome P450; drug-drug interaction; molecular docking; pharmacokinetics.

FIGURE 1

(A) Michaelis-Menten plots of enzymatic activity of RLMs, HLMs and rCYP3A4 on tofacitinib metabolism. Data are presented as the mean ± standard deviation of three experiments. (B) Screening of bergapten, psoralidin, xanthotoxol, psoralen, isopsoralen, and 8-methoxypsoralen in RLMs. Values are mean ± SD, n = 3. *p < 0.05, significant difference from control group.

FIGURE 2

(A) IC₅₀ of the inhibition of tofacitinib in RLMs by bergapten; (B) IC₅₀ of the inhibition of tofacitinib in HLMs by bergapten; (C) IC₅₀ of the inhibition of tofacitinib in rCYP3A4 by bergapten. Values are mean ± SD, n = 3.

FIGURE 3

Inhibitory mechanism of bergapten against tofacitinib in (A,B) RLMs, (C,D) HLMs and (E,F) rCYP3A4. Values are mean ± SD, n = 3.

FIGURE 4

(A) IC₅₀ of tofacitinib inhibition in RLMs by isopsoralen; (B) IC₅₀ of tofacitinib inhibition in HLMs by isopsoralen; (C) IC₅₀ of tofacitinib inhibition in rCYP3A4 by isopsoralen.

FIGURE 5

Inhibitory mechanism of isopsoralen against tofacitinib in (A,B) RLMs, (C,D) HLMs and (E,F) rCYP3A4. Values are mean ± SD, n = 3.

FIGURE 6

Molecular docking scheme of tofacitub and bergapten (A) and isopsoralen (C); Action site between bergapten (B), isopsoralen (D) and tofacitinib and CYP3A4 via hydrogen bonding.

FIGURE 7

Mean plasma concentration–time curves of tofacitinib in different treatment groups. Group (A), bergapten; group (B), isopsoralen (n = 6).