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Published on: August 27, 2015
Metabolic profiling in liver microsomes and mice of E28, a potent FLT3 inhibitor
Yan Tan1, Xiandeng Li1, Minghai Tang2
1School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, China.
Abstract:
The objective of this study was to clarify the species differences of metabolic stability of E28 in liver microsomes, and to study metabolic phenotypes of E28 in human liver microsomes by chemical inhibition method.The metabolites in plasma, urine, and faeces samples from mice received caudal vein intravenous were detected and identified by UHPLC-HRMS, and the tissue distribution was studied after oral administration.E28 was metabolised rapidly in liver microsomes of each species with a short half-live T1/2 and a moderate clearance, except for rats. The metabolic properties of E28 were similar in human and mouse liver microsomes. Data from metabolic phenotype studies indicated that CYP2D6, CYP3A4 and CYP2C9 were the main metabolic enzymes participating in the metabolism of E28.The main metabolic pathways implicated include oxidation, methylation, amide hydrolysis, acetylation, glucuronide conjugation.Tissue distribution studies showed that E28 could be detected in all organs and tissues after oral administration, with the highest level in the stomach and the lowest in the brain. In bone marrow cells, the concentration of E28 in all sample points were consistently higher than its half inhibitory concentration against MV4-11 tumour cells.
Insights
E28 is rapidly metabolized in liver microsomes across species, with similar metabolic profiles in humans and mice. Key enzymes like CYP2D6, CYP3A4, and CYP2C9 are involved in its breakdown.
Area of Science:
- Pharmacokinetics and Drug Metabolism
- Biochemistry
- Toxicology
Background:
- Understanding drug metabolism is crucial for predicting efficacy and safety.
- Species differences in drug metabolism can impact preclinical to clinical translation.
- E28's metabolic stability and pathways require detailed investigation.
Purpose of the Study:
- To elucidate species-specific metabolic stability of E28 in liver microsomes.
- To determine the metabolic phenotype of E28 in human liver microsomes using chemical inhibition.
- To characterize the tissue distribution and major metabolic pathways of E28.
Main Methods:
- In vitro metabolic stability assays using liver microsomes from various species.
- Chemical inhibition studies with specific cytochrome P450 enzymes (CYPs).
- Ultra-High-Performance Liquid Chromatography-High-Resolution Mass Spectrometry (UHPLC-HRMS) for metabolite identification.
- Tissue distribution studies following oral administration in mice.
Main Results:
- E28 exhibited rapid metabolism in liver microsomes across species, except rats, with a short half-life and moderate clearance.
- Metabolic properties were comparable between human and mouse liver microsomes.
- CYP2D6, CYP3A4, and CYP2C9 were identified as the primary enzymes responsible for E28 metabolism.
- Identified metabolic pathways included oxidation, methylation, amide hydrolysis, acetylation, and glucuronide conjugation.
- E28 distributed to all tissues, with highest concentrations in the stomach and lowest in the brain.
- Concentrations in bone marrow cells exceeded the half inhibitory concentration against MV4-11 tumor cells.
Conclusions:
- E28 undergoes rapid metabolism, primarily mediated by CYP2D6, CYP3A4, and CYP2C9.
- Human and mouse liver microsomes provide relevant models for studying E28 metabolism.
- The tissue distribution suggests potential accumulation in certain organs, with notable levels in bone marrow.
- Further investigation into E28's pharmacological activity and safety profile is warranted based on its metabolic and distribution characteristics.
