Application of newly developed and validated UPLC-MS/MS method for pharmacokinetic study of ROS1/NTRK inhibitor
Yunfeng Zhu1, Fangkai Wang1, Xin Wang1
1Department of Emergency Surgery, The First Affiliated Hospital of Anhui Medical University, Hefei 230022, Anhui Province, China.
Abstract:
Taletrectinib is a potent selective ROS and pan-NTRK tyrosine kinase inhibitor (TKI) and has been developed to treat non-small cell lung cancer (NSCLC). To facilitate pharmacokinetic and toxicokinetic studies of taletrectinib, we developed a procedure for ultra-high-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) to detect the plasma level of taletrectinib in dogs. This assay procedure was validated in compliance with FDA guidance. The dog plasma samples were spiked with internal standard (IS), followed by protein precipitation, and analyzed using a Waters ACQUITY BEH C18 column coupled to a Thermo triple quadrupole mass spectrometer. Separation was executed using the acetonitrile-0.1 % formic acid solution with gradient elution, at a flow rate of 0.4 mL/min. Taletrectinib and IS were monitored by multiple reaction monitoring (MRM) with m/z 406.2 > 349.2 and m/z 441.2 > 138.1, respectively. The procedure demonstrated excellent linearity with a correlation coefficient greater than 0.999 within the concentration range of 0.2-200 ng/mL. The inter- and intra-day accuracy ranged from -5.25 % to 5.26 %, and the precision was below 6.39 %. Acetonitrile-mediated protein precipitation showed high extraction efficiency and a recovery above 85 %. The procedure was then applied to quantify taletrectinib in beagle dog plasma after oral and intravenous doses and achieved success. The obtained pharmacokinetic parameters indicated high bioavailability of taletrectinib (>85 %) and extensive tissue distribution (>40 L/kg).
Insights
A validated UPLC-MS/MS method accurately measures taletrectinib in dog plasma, crucial for non-small cell lung cancer drug development. This assay reveals high bioavailability and extensive tissue distribution for taletrectinib.
Area of Science:
- Pharmacology
- Analytical Chemistry
- Oncology
Background:
- Taletrectinib is a tyrosine kinase inhibitor (TKI) targeting ROS and NTRK, developed for non-small cell lung cancer (NSCLC).
- Pharmacokinetic and toxicokinetic studies are essential for drug development, requiring accurate plasma concentration measurements.
Purpose of the Study:
- To develop and validate an ultra-high-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for quantifying taletrectinib in dog plasma.
- To support pharmacokinetic and toxicokinetic assessments of taletrectinib.
Main Methods:
- Plasma samples were prepared using protein precipitation with acetonitrile and an internal standard.
- Analysis was performed on a UPLC system coupled to a triple quadrupole mass spectrometer using gradient elution.
- Taletrectinib and internal standard were detected via multiple reaction monitoring (MRM).
Main Results:
- The UPLC-MS/MS method exhibited excellent linearity (R² > 0.999) over a 0.2-200 ng/mL range.
- Assay accuracy ranged from -5.25% to 5.26%, with precision below 6.39%.
- Extraction recovery was consistently above 85%, and the method was successfully applied to quantify taletrectinib in vivo.
Conclusions:
- A validated UPLC-MS/MS assay provides a reliable method for measuring taletrectinib in dog plasma.
- Pharmacokinetic studies in dogs demonstrated high taletrectinib bioavailability (>85%) and extensive tissue distribution (>40 L/kg).
- This validated assay is critical for advancing taletrectinib's development for NSCLC treatment.
