Quantitation of Tuvusertib (M1774) in Human Plasma by LC-MS/MS
Tien V Le1,2, Robert A Parise1, Julianne L Holleran1
1Cancer Therapeutics Program, UPMC Hillman Cancer Center, Pittsburgh, Pennsylvania, USA.
Abstract:
Ataxia-telangiectasia and Rad3-related (ATR) protein kinase is an essential regulator of the DNA damage response (DDR) at stalled and collapsed replication forks. Tuvusertib (M1774) is a selective, orally available small molecule ATR inhibitor currently in preclinical and clinical development for cancer treatment. This study presents a robust and simple 5-min assay designed for the quantification of single agent tuvusertib in human plasma utilizing liquid chromatography tandem mass spectrometry (LC-MS/MS). A 20 μL volume of plasma was subjected to protein precipitation, followed by chromatographic separation using a Phenomenex Synergi Polar-RP (4 μm, 2.1 × 50 mm) and a gradient mobile phase system consisting of 0.1% formic acid in both water and acetonitrile during a 4-min run time. Mass spectrometric detection was achieved using a SCIEX 6500+ tandem mass spectrometer with electrospray positive-mode ionization. With a stable isotopic internal standard, our assay met the criteria outlined by the Food and Drug Administration guidance for bioanalytical method validation, demonstrating robust performance within the range from 5 to 5000 ng/mL. This assay will support ongoing and future clinical studies by defining tuvusertib pharmacokinetics.
Insights
A new 5-minute assay quantifies tuvusertib, an ATR inhibitor, in human plasma using LC-MS/MS. This validated method supports clinical trials by measuring tuvusertib pharmacokinetics for cancer treatment.
Area of Science:
- Pharmacology
- Analytical Chemistry
- Cancer Biology
Background:
- Ataxia-telangiectasia and Rad3-related (ATR) protein kinase is crucial for DNA damage response.
- Tuvusertib (M1774) is an ATR inhibitor in clinical development for cancer therapy.
Purpose of the Study:
- To develop and validate a rapid and robust bioanalytical assay for quantifying tuvusertib in human plasma.
- To support clinical studies by enabling accurate pharmacokinetic analysis of tuvusertib.
Main Methods:
- Liquid chromatography tandem mass spectrometry (LC-MS/MS) was employed for quantification.
- A 5-minute assay involving protein precipitation and gradient elution on a Phenomenex Synergi Polar-RP column was utilized.
- Detection was performed on a SCIEX 6500+ mass spectrometer using positive-mode electrospray ionization.
Main Results:
- The assay demonstrated robust performance and met FDA bioanalytical method validation criteria.
- Quantification was achieved within a wide dynamic range of 5 to 5000 ng/mL.
- A stable isotopic internal standard was used for enhanced accuracy.
Conclusions:
- A validated 5-minute LC-MS/MS assay for tuvusertib in human plasma has been established.
- This assay is suitable for supporting pharmacokinetic studies in ongoing and future clinical trials of tuvusertib.
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