= Discovery stage. (24.37%, 2023)
= Translation stage. (39.50%, 2023)
= Clinically available. (36.13%, 2023)
MSACL 2023 : Kirkpatrick

MSACL 2023 Abstract

Self-Classified Topic Area(s): Tox / TDM / Endocrine > Precision Medicine > Assays Leveraging MS

Poster Presentation
Poster #47a
Attended on Thursday at 11:00

Method Development and Validation of Paper Spray Mass Spectrometry Method for Quantitation of Remdesivir and Active Metabolite, GS-441524, in Plasma

Hannah Zimmerman-Federle (1), Christine Skaggs (1), Nicholas Manicke (1, 2), Lindsey Kirkpatrick (3)
(1) Department of Chemistry and Chemical Biology, Indiana University-Purdue University Indianapolis, Indianapolis, IN 46202, USA.(2) Forensics and Investigative Sciences, Indiana University-Purdue University Indianapolis, Indianapolis, IN 46202, USA.(3) Department of Pediatrics, Division of Pediatric Infectious Diseases, Indiana University School of Medicine, Indianapolis, IN 46202, USA.

Lindsey Kirkpatrick, DO, PhD (Presenter)
Indiana University School of Medicine

Abstract

Introduction:
Remdesivir (GS-5734) is a nucleoside analog prodrug with antiviral activity against several single-stranded RNA viruses, including ebolavirus (EBOV), severe respiratory distress syndrome virus 1 (SARS-CoV-1), and Middle East respiratory syndrome coronavirus (MERS-CoV). In vitro studies of remdesivir and its active metabolite, GS-441524, also showed antiviral activity against the novel severe respiratory distress syndrome virus 2 (SARS-CoV-2), the causative viral pathogen of COVID-19, and it is one of the FDA-approved antiviral agents for the treatment of individuals with COVID-19. However, remdesivir pharmacokinetics and pharmacodynamics (PK/PD) data in humans is limited. It is imperative that precise analytical methods for the quantification of remdesivir and GS-441524 for use in PK/PD studies, therapeutic drug monitoring, and assessment of toxicity are developed.

Methods:
Seven-point calibration curves for remdesivir and its active metabolite, GS-441524, were created utilizing seven plasma-based calibrants of varying concentrations and two isotopic internal standards of set concentrations. Four plasma-based quality controls were prepared in a similar fashion to the calibrants and utilized for validation. No sample preparation was needed. Plasma samples were spotted on a paper substrate in pre-manufactured Verispray plastic cassettes and allowed to dry. The dried plasma spots were analyzed directly utilizing paper spray-mass spectrometry (PS-MS/MS). The method was validated, and the success of the validation was assessed by evaluating linearity, limits of detection (LOD), accuracy (% bias), precision (% CV), carryover, matrix effects, stability, and metabolic interferences. All experiments were performed on a Thermo Scientific Altis triple quadrupole mass spectrometer.

Results:
The calibration ranges were 20 – 5,000 and 100 – 25,000 ng/mL for remdesivir and GS-441524, respectively. The calibration curves for the two antiviral agents showed excellent linearity (average R2 = 0.99-1.00). The inter- and intra-day precision (%CV) across validation runs at four QC levels for both analytes was less than 11.2% and accuracy (%bias) was within ± 15%. Plasma calibrant stability was assessed and degradation for the 4 °C and room temperature samples were seen beginning at Day 7. The plasma calibrants were stable at -20 °C. No interference, matrix effects, or carryover was discovered during the validation process. The final method had an analysis time of 1.2 minutes.

Conclusion:
The development and validation of the first PS-MS/MS method quantitating remdesivir and its active metabolite, GS-441524, is reported. PS-MS/MS represents a useful methodology for rapidly quantifying remdesivir, which will be useful for clinical PK/PD, therapeutic drug monitoring, and toxicity assessment, particularly during future viral outbreaks.


Financial Disclosure

DescriptionY/NSource
GrantsyesNIH, Miravista Laboratories
Salaryno
Board Memberno
Stockno
Expensesno
IP Royaltyno

Planning to mention or discuss specific products or technology of the company(ies) listed above:

no