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Optimization of a sensitive and reliable UPLC-MS/MS method to simultaneously quantify almonertinib and HAS-719 and
Dongxin Chen1, Jie Chen2, Yuxin Shen2
1The Affiliated Lihuili Hospital, Ningbo University, Ningbo, Zhejiang, China.
Context:
Almonertinib is primarily metabolized by CYP3A4, so it could interact with a variety of drugs metabolized by CYP3A4, leading to the changes of systemic exposure.
Objective:
For the purpose of this experiment, an ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) assay with accuracy and simplicity was optimized and fully validated for the simultaneous quantitative determination of almonertinib and its metabolite HAS-719, and drug-drug interactions (DDI) between almonertinib and nicardipine in vivo and in vitro was researched.
Materials And Methods:
Detection of analytes was achieved by UPLC-MS/MS coupled with multiple reaction monitoring (MRM) in the positive ion mode with ion transitions of m/z 526.01 → 72.04 for almonertinib, m/z 512.18 → 455.08 for HAS-719 and m/z 447.16 → 128.11 for IS, respectively.
Results:
There was favourable linearity in the 0.5-200 ng/mL calibration range for almonertinib and 0.5-100 ng/mL for HAS-719. The lower limit of quantification (LLOQ) for both analytes was 0.5 ng/mL. The precision, accuracy, stability, matrix effect and extraction recovery required for methodological validation were consistent with the requirements of FDA guideline. Then, the UPLC-MS/MS assay was employed successfully on the interactions of almonertinib and nicardipine in vivo and in vitro. The half-maximal inhibitory concentration (IC50) was 1.19 μM in rat liver microsomes (RLM), where nicardipine inhibited the metabolism of almonertinib with a mixed inhibitory mechanism. In pharmacokinetic experiments of rats, it was observed that nicardipine could significantly alter the pharmacokinetic profiles of almonertinib, including AUC(0-∞), AUC(0-t) and Cmax, but had no effect on the metabolism of HAS-719.
Conclusion:
According to the findings, it was indicated that nicardipine could inhibit the metabolism of almonertinib in vitro and in vivo.
Insights
Nicardipine inhibits almonertinib metabolism, altering its pharmacokinetic profiles. This study developed a validated UPLC-MS/MS assay to quantify almonertinib and its metabolite, confirming drug-drug interactions in vitro and in vivo.
Area of Science:
- Pharmacology
- Drug Metabolism
- Analytical Chemistry
Background:
- Almonertinib is metabolized by CYP3A4, suggesting potential interactions with other CYP3A4 substrates.
- Drug-drug interactions (DDIs) can alter drug exposure and efficacy.
Purpose of the Study:
- To develop and validate a UPLC-MS/MS assay for quantifying almonertinib and its metabolite HAS-719.
- To investigate the in vitro and in vivo drug-drug interactions between almonertinib and nicardipine.
Main Methods:
- An ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) assay was optimized and validated.
- Simultaneous determination of almonertinib and HAS-719 using multiple reaction monitoring (MRM).
- In vitro studies using rat liver microsomes and in vivo pharmacokinetic studies in rats.
Main Results:
- The UPLC-MS/MS assay demonstrated linearity, accuracy, and precision within FDA guidelines.
- Nicardipine inhibited almonertinib metabolism in rat liver microsomes with a mixed mechanism (IC50 = 1.19 μM).
- Nicardipine significantly altered almonertinib's pharmacokinetic parameters (AUC, Cmax) in rats but did not affect HAS-719 metabolism.
Conclusions:
- Nicardipine inhibits almonertinib metabolism both in vitro and in vivo.
- The validated UPLC-MS/MS assay is suitable for studying almonertinib pharmacokinetics and DDIs.
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