Sorafenib and Sunitinib, Two Anticancer Drugs, Inhibit CYP3A4-Mediated and Activate CY3A5-Mediated Midazolam 1′-Hydroxylation

Abstract

ABSTRACT:Sorafenib and sunitinib are novel small-molecule molecularly targeted anticancer drugs that inhibit multiple tyrosine kinases. These medicines have shown survival benefits in advanced renal cell carcinomas as well as in advanced hepatocellular carcinomas and gastrointestinal stromal tumors, respectively. The effects of sorafenib and sunitinib on midazolam 1-hydroxylation catalyzed by human CYP3A4 or CYP3A5 were investigated. Sorafenib and sunitinib inhibited metabolic reactions catalyzed by recombinant CYP3A4. Midazolam hydroxylation was also inhibited in human liver microsomes harboring the CYP3A5*3/*3 genotype (poor CYP3A5 expressor). In contrast, midazolam 1-hydroxylation catalyzed by recombinant CYP3A5 was enhanced by the coexistence of sorafenib or sunitinib in a concentration-dependent manner, with saturation occurring at approximately 10 M. Midazolam hydroxylation was also enhanced in human liver microsomal samples harboring the CYP3A5*1/*1 genotype (extensive CYP3A5 expressor).Sorafenib N-oxidation and sunitinib N-deethylation, the primary routes of metabolism, were predominantly catalyzed by CYP3A4 but not by CYP3A5. The preincubation period of sorafenib and sunitinib before the midazolam addition in the reaction mixture did not affect the enhancement of CYP3A5-catalyzed midazolam hydroxylation, indicating that the enhancement was caused by parent sorafenib and sunitinib. Docking studies with a CYP3A5 homology model based on the structure of CYP3A4 revealed that midazolam closely docked to the heme of CYP3A5 compared with sorafenib or sunitinib, suggesting that these anticancer drugs act as enhancers, not as substrates. Our results thus showed that sorafenib and sunitinib activated midazolam 1-hydroxylation by CYP3A5 but inhibited that by CYP3A4. Unexpected drug interactions involving sorafenib and sunitinib might occur via heterotropic cooperativity of CYP3A5.

In Vitro Assessment of Time-Dependent Inhibitory Effects on CYP2C8 and CYP3A Activity by Fourteen Protein Kinase Inhibitors

“…Sorafenib and sunitinib have also been reported to activate the formation of 19-hydroxymidazolam (Sugiyama et al, 2011;Kenny et al, 2012), but this was not observed in our incubations. The heteroactivation mechanism for these kinase inhibitors has been reported to be substratedependent, as it has been observed with midazolam but not with the other CYP3A marker substrates nifedipine and testosterone (Li et al, 2007;Dong et al, 2011;Kenny et al, 2012).…”

“…The heteroactivation mechanism for these kinase inhibitors has been reported to be substratedependent, as it has been observed with midazolam but not with the other CYP3A marker substrates nifedipine and testosterone (Li et al, 2007;Dong et al, 2011;Kenny et al, 2012). It is not clear whether the activation by erlotinib and gefitinib occurs via CYP3A4 or CYP3A5, but for sorafenib and sunitinib it is reported to occur via CYP3A5 (Sugiyama et al, 2011). In the present study, we did not differentiate between CYP3A4 and CYP3A5, but the lack of stimulation of midazolam 19-hydroxylation by sorafenib and sunitinib suggests that CYP3A4 activity was more prominent than that of CYP3A5 in the HLM batch used.…”

CYP3A4 overexpression enhances apoptosis induced by anticancer agent imidazoacridinone C-1311, but does not change the metabolism of C-1311 in CHO cells

“…The latter strongly inhibited CYP3A4 while also being metabolized by it. However, there are few reports about the inhibition of specific P450 isoenzymes by antitumor agents, eg, by thiotepa [42,43] , tamoxifen [44,45] and the less well-known sorafenib [46] . Our results showed that cytochrome P450 CYP3A4 did not metabolize C-1311 in cells, confirming our hypothesis that overexpression of this enzyme might modulate the cellular response of CHO cells following C-1311 treatment, even though CYP3A4 does not participate in metabolic transformations.…”

Drug Interaction of Efavirenz and Midazolam: Efavirenz Activates the CYP3A-Mediated Midazolam 1′-Hydroxylation In Vitro

“…Heterotropic positive cooperativity may also explain the interaction of thalidomide with midazolam in human CYP3A5 in which thalidomide increased midazolam 1Ј-hydroxylation and total midazolam oxidation (Okada et al, 2009). Midazolam 1Ј-hydroxylation was shown to be activated by sorafenib and sunitinib in CYP3A5 (Sugiyama et al, 2011) and ticagrelor in HLM (Zhou et al, 2011).…”