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A novel uHPLC-MS/MS method for the quantitation of AZD7451 (AZ12607092) in human plasma
Cody J Peer1, Jeffrey L Brown, Timothy J Martin
1Clinical Pharmacology Program, Office of the Clinical Director, National Cancer Institute, Bethesda, MD, United States.
Abstract:
Tropomyosin-related kinases (Trk) are tyrosine kinase receptors implicated in tumor proliferation, invasion, and survival signaling across a number of tumors, making them potentially attractive targets for the treatment of cancer. AZD7451 is a potent and selective inhibitor of Trk kinases currently undergoing a Phase I dose escalation in glioblastoma multiforme at the National Cancer Institute. A key part of early clinical testing for AZD7451 involves demonstrating that pharmacokinetic half-life and clinical exposures of AZD7451 are sufficient to inhibit Trk receptors in preclinical models. To address this need, an ultra sensitive analytical method was developed to measure the AZD7451 profile in human plasma. A liquid-liquid extraction recovered >80% of AZD7451 before quantitative analysis by ultra HPLC-MS/MS. A Varian Polaris(®) C18-A column and a mass transition of m/z 383.5→340.5 (m/z 389.6→342.0 for the internal standard [(2)H6]-AZD7451) was used, and a dynamic calibration range of 0.5-1000ng/mL was established, which provided a sensitive (<8.5% deviation), and precise (<6%) quantitative assay for AZD7451. AZD7451 demonstrated stability in human plasma at room temperature for 24h (<7% change) and after extraction at 4°C for 24h (<8% change), and was stable through 4 freeze/thaw cycles (<8% change). This method was used to measure AZD7451 plasma levels in clinical samples to confirm the sensitivity at several time points following AZD7451 treatment in subjects with glioblastoma.
Insights
A new ultra-sensitive method accurately measures AZD7451 in human plasma, crucial for cancer drug development. This assay supports clinical trials by confirming sufficient drug exposure for Tropomyosin-related kinases (Trk) inhibition in glioblastoma patients.
Area of Science:
- Oncology
- Pharmacology
- Analytical Chemistry
Background:
- Tropomyosin-related kinases (Trk) are key drivers in cancer proliferation and survival.
- AZD7451 is a selective Trk inhibitor in Phase I trials for glioblastoma multiforme.
- Accurate measurement of drug levels is essential for assessing efficacy in early clinical trials.
Purpose of the Study:
- To develop and validate an ultra-sensitive analytical method for quantifying AZD7451 in human plasma.
- To establish the pharmacokinetic profile of AZD7451 in glioblastoma patients.
- To ensure sufficient drug exposure for Trk receptor inhibition.
Main Methods:
- Liquid-liquid extraction followed by ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS).
- Utilized a Varian Polaris C18-A column with a specific mass transition for AZD7451 and its deuterated internal standard.
- Established a dynamic calibration range of 0.5-1000 ng/mL with high sensitivity and precision.
Main Results:
- The method demonstrated high sensitivity (<8.5% deviation) and precision (<6%).
- AZD7451 exhibited stability in human plasma under various conditions (room temperature, 4°C, freeze-thaw cycles).
- The assay successfully measured AZD7451 plasma levels in clinical samples from glioblastoma subjects.
Conclusions:
- An ultra-sensitive and robust UHPLC-MS/MS method was successfully developed for AZD7451 quantification in human plasma.
- The method supports early clinical testing by confirming adequate drug exposure for Trk inhibition.
- This assay is vital for ongoing Phase I trials of AZD7451 in glioblastoma.
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