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Bioanalysis of alectinib and metabolite M4 in human plasma, cross-validation and impact on PK assessment
Katja Heinig1, Kazuhiro Miya2, Tomonori Kamei2
1Roche Pharmaceutical Research & Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, 4070 Basel, Switzerland.
Background:
Alectinib is a novel anaplastic lymphoma kinase (ALK) inhibitor for treatment of patients with ALK-positive non-small-cell lung cancer who have progressed on or are intolerant to crizotinib. To support clinical development, concentrations of alectinib and metabolite M4 were determined in plasma from patients and healthy subjects.
Methods:
LC-MS/MS methods were developed and validated in two different laboratories: Chugai used separate assays for alectinib and M4 in a pivotal Phase I/II study while Roche established a simultaneous assay for both analytes for another pivotal study and all other studies.
Conclusion:
Cross-validation assessment revealed a bias between the two bioanalytical laboratories, which was confirmed with the clinical PK data between both pivotal studies using the different bioanalytical methods.
Insights
Bioanalytical methods for alectinib and its metabolite M4 showed bias between laboratories. This impacted pharmacokinetic data in patients with anaplastic lymphoma kinase-positive non-small-cell lung cancer.
Area of Science:
- Pharmacology
- Analytical Chemistry
- Oncology
Background:
- Alectinib is an anaplastic lymphoma kinase (ALK) inhibitor for ALK-positive non-small-cell lung cancer (NSCLC) patients progressing on or intolerant to crizotinib.
- Accurate quantification of alectinib and its metabolite M4 in plasma is crucial for clinical development.
Purpose of the Study:
- To develop and validate bioanalytical methods for quantifying alectinib and M4 in human plasma.
- To assess the consistency of these methods across different laboratories.
Main Methods:
- Liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods were developed and validated.
- Two laboratories (Chugai and Roche) used distinct assay strategies: separate assays for alectinib and M4 versus a simultaneous assay for both analytes.
- Methods were applied to plasma samples from patients and healthy subjects.
Main Results:
- Cross-validation between the two laboratories revealed a significant bias in analyte quantification.
- This bias was confirmed by analyzing clinical pharmacokinetic (PK) data from pivotal studies.
Conclusions:
- Discrepancies in bioanalytical methods between laboratories can introduce bias into drug concentration measurements.
- Careful method validation and cross-validation are essential to ensure reliable PK data for drug development.
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