Crizotinib inhibits the metabolism of tramadol by non-competitive suppressing the activities of CYP2D1 and CYP3A2

Abstract

Objectives: To investigate the interaction between tramadol and representative tyrosine kinase inhibitors, and to study the inhibition mode of drug-interaction.

Methods: Liver microsomal catalyzing assay was developed. Sprague-Dawley rats were administrated tramadol with or without selected tyrosine kinase inhibitors. Samples were prepared and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used for analysis. Besides, liver, kidney, and small intestine were collected and morphology was examined by hematoxyline-eosin (H&E) staining. Meanwhile, liver microsomes were prepared and carbon monoxide differential ultraviolet radiation (UV) spectrophotometric quantification was performed.

Results: Among the screened inhibitors, crizotinib takes the highest potency in suppressing the metabolism of tramadol in rat/human liver microsome, following non-competitive inhibitory mechanism. In vivo, when crizotinib was co-administered, the AUC value of tramadol increased compared with the control group. Besides, no obvious pathological changes were observed, including cell morphology, size, arrangement, nuclear morphology with the levels of alanine transaminase (ALT) and aspartate transaminase (AST) increased after multiple administration of crizotinib. Meanwhile, the activities of CYP2D1 and CYP3A2 as well as the total cytochrome P450 abundance were found to be decreased in rat liver of combinational group.

Conclusions: Crizotinib can inhibit the metabolism of tramadol. Therefore, this recipe should be vigilant to prevent adverse reactions.

Keywords: CYP; Combination; Crizotinib; Non-competitive; Tramadol.

Figure 1. Determine the interaction between representative tyrocine kinase inhibitors and tramadol.

(A) The inhibitory effect of 23 types of tyrosine kinase inhibitors on the production of O-desmethyl tramadol in RLM compared with the control group. (B) Evaluate the half-maximal inhibitory concentration (IC₅₀) of crizotinib with various concentrations (0.01, 0.1, 1, 10, 25, 50 and 100 µM) on tramadol metabolism in RLM and HLM. Data are presented as the mean ± SD, n = 3.

Figure 2. Primary lineweaver-Burk plot and secondary plot for K_i and αK_i in the inhibition of tramadol metabolism by crizotinib with various concentrations in (A) RLM and (B) HLM, respectively.

Data are presented as the mean ± SD, n = 3.

Figure 3. Mean concentration–time curve of tramadol and O-desmethyl tramadol in four groups.

(A) Evaluate the effect of a single dose (45 mg/kg) of crizotinib. (B) Evaluate the effect of multiple doses (45 mg/kg) of crizotinib for 7 days. Data are presented as the mean ± SD, n = 5.

Figure 4. The effect of crizotinib on tissue morphology and liver function.

(A) The results of H&E staining of tissue sections for liver (a), kidney (b) and small intestine (c) in three groups. (B) The value of serum ALT (U/L) and AST (U/L) in three groups. Data are presented as the mean ± SD, n = 3; Vs Control, * P < 0.05, ** P < 0.01.

Figure 5. Crizotinib suppressing the activities of CYP by reducing the abundance of CYP enzymes.

(A) The effect of crizotinib on the function of CYP2D1 (dextromethorphan as probe substrate) and CYP3A2 (midazolam as probe substrate) in the liver of three groups. (B) The chromatogram and (C) the total amount of CYP in rat liver microsomes quantified by CO quantitative method in three groups. Data are presented as the mean ± SD, n = 3; Vs Control, *** P < 0.001.