Identification of Novel Pathways of Osimertinib Disposition and Potential Implications for the Outcome of Lung Cancer Therapy

Abstract

2 Abstract PurposeOsimertinib is a third-generation inhibitor of the epidermal growth factor receptor used in treatment of non-small cell lung cancer. A full understanding of its disposition and capacity for interaction with other medications will facilitate its effective use as a single agent and in combination therapy. Experimental designRecombinant cytochrome P450s and liver microsomal preparations were used to identify novel pathways of osimertinib metabolism in vitro. A panel of knockout and mouse lines humanized for pathways of drug metabolism were used to establish the relevance of these pathways in vivo. ResultsAlthough some osimertinib metabolites were similar in mouse and human liver samples there were several significant differences, in particular a marked species difference in the P450s involved. The murine Cyp2d gene cluster played a predominant role in mouse, whereas CYP3A4 was the major human enzyme responsible for osimertinib metabolism. Induction of this enzyme in CYP3A4 humanized mice substantially decreased circulating osimertinib exposure. Importantly, we discovered a further novel pathway of osimertinib disposition involving CPY1A1. Modulation of CYP1A1/CYP1A2 levels markedly reduced parent drug concentrations, significantly altering metabolite pharmacokinetics (PK) in humanized mice in vivo. ConclusionWe demonstrate that a P450 enzyme expressed in smokers' lungs and lung tumors has the capacity to metabolise osimertinib. This could be a significant factor in defining the outcome of osimertinib treatment. This work also illustrates how P450-humanized mice can be used to identify and mitigate Acquired resistance to first and second-generation EGFR inhibitors usually involves a further EGFR mutation, T790M. Osimertinib (AZD9291) is the most effective treatment in these latter cases and, due to its low toxicity and high level of brain penetration, is currently being explored as a first-line therapy for metastatic disease. Osimertinib is a cytochrome P450 substrate. These enzymes can define both systemic exposure and intra-tumoral drug concentrations and can therefore be major determinants in the outcome of cancer therapy. We report a novel pathway of osimertinib disposition which may be of importance in this regard.Research.on May 9, 2018.

An Extensively Humanized Mouse Model to Predict Pathways of Drug Disposition and Drug/Drug Interactions, and to Facilitate Design of Clinical Trials

“…4). These data, consistent with our previous report using a simpler humanized model (MacLeod et al, 2018), suggest that members of the CYP1A gene family are involved in osimertinib disposition. The major human metabolites of osimertinib were detected in 8HUM mice (Fig.…”

<p>Metabolite Profiling in Anticancer Drug Development: A Systematic Review</p>

“…Out of 78 articles included in the analysis (42 in vitro, 42 in vivo, and 16 clinical studies), 47 (60.3%), 14 (17.9%) and 17 (21.8%) articles respectively, investigated metabolite profiles of conventional anticancer drugs/synthetic anticancer candidates, small-molecules targeted therapy, and herbderived compounds with anticancer activities. These studies involved a total of 57 (57.0%) conventional anticancer drugs/ synthetic anticancer candidates, 6, 22 (22.0%) studies for small-molecules targeted therapy, 4,5,[59][60][61][62][63][64][65][66][67][68][69][70] and 21 (21.0%) studies for herb-derived compounds with anticancer activities 2,3,7,9,[71][72][73][74][75][76][77][78][79][80][81][82][83] (Tables 1-3). Anticancer drugs or candidate compounds are metabolized by either Phase I, or phase II metabolizing enzyme alone, or both phase I and phase II metabolizing enzymes.…”

“…Antitumor (breast carcinoma), Apoptotic induction SN30000 (3-(3-Morpholinopropyl)-7,8-dihydro-6H-indeno[5,6-e] [1,2,4] triazine 1.4-dioxide: tirapazamine analog)[6,94] Antiproliferative (Human colon cancer (HT29) and cervical (SiHa) cancer cells), Antitumor (HT29 and SiHa cancer cells models)Tamoxifen[54,55] Antiestrogen(Breast cancer) TAS-102 (tipiracil hydrochloride (TPI) + trifluridine (FTD)) [56] Anticancer (Metastatic colorectal cancer) TNP-470 (O-(chloroacetyl-carbamoyl) fumagillol) [57] Antitumor (B16 BL6 melanoma, M 5076 reticulum cell sarcoma, Lewis lung carcinoma, Walker 256 carcinoma), Anticancer, Antiangiogenesis TW01003 ((3E,6E)-3-Benzylidene-6-[(5-hydroxypyridin-2-yl) methylene] piperazine-2,5-dione) induction, and Migration reduction (Leukemia, cervical, breast cancer, and laryngeal carcinoma) Brivanib [60,95] Anticancer (Hepatocellular carcinoma) EPZ-5676 (pinometostat) [61] Anticancer (Leukemia) HM781-36B (1-[4-[4-(3,4-dichloro-2-fluorophenylamino)-7methoxyquinazolin-6-yloxy]-piperidin-1-yl] prop-2-en-1-one hydrochloride) [5] Anticancer (Advanced solid tumors i.e. non-small-cell lung cancer (NSCLC) patients with EGFR mutation (T790M), breast, , non-small-cell lung, and gastric cancer and glioblastoma cells models) Lenvatinib [66] Anticancer (Hepatocellular carcinoma, melanoma, renal carcinoma, non-small-cell lung carcinoma, glioblastoma multiforme, ovarian and endometrial carcinoma)Neratinib[67] Anticancer (Breast cancer) ON 013100 ((E)-2,4,6-trimethoxystyryl-3-hydroxy-4methoxybenzyl sulfone (Kinase inhibitor))[68] Anticancer (Lymphoma and acute lymphoid leukemia) Osimertinib[69] Anticancer (Non-small cell lung cancer) Pimasertib[70] Anticancer (Non-small cell lung, colorectal cancer, and head and neck squamous cell carcinoma) TKI258 (dovitinib)[96] Anticancer (Renal cell carcinoma, advanced breast cancer, relapsed multiple myeloma and urothelial cancer)…”

P450-Humanized and Human Liver Chimeric Mouse Models for Studying Xenobiotic Metabolism and Toxicity

“…Although the necessity to preinduce transgene expression with a CYP1A inducer [2,3,7,] makes the study complicated to conduct and interpret, the fact that human CYP1A-specific metabolites are formed in vivo illustrated the potential utility of the model for in vivo drug metabolism studies. Two recent articles (Lin et al, 2017;MacLeod et al, 2018) also described the utility of a newly generated CYP1A1/CYP1A2-humanized model in studying the disposition of anticancer drugs; however, details regarding the mouse model were not provided.…”