Document Type
Article
Publication Date
5-15-2023
Abstract
Rationale
There is currently no treatment for spinocerebellar ataxias (SCAs), which are a group of genetic disorders that often cause a lack of coordination, difficulty walking, slurred speech, tremors, and eventually death. Activation of KCa2.2/KCa2.3 channels reportedly exerts beneficial effects in SCAs. Here, we report the development and validation of an analytical method for quantitating a recently-developed positive allosteric modulator of KCa2.2/KCa2.3 channels (compound 2q) in mouse plasma.
Methods
Mouse plasma samples (10 μL) containing various concentrations of 2q were subjected to protein precipitation in the presence of a structurally similar internal standard (IS). Subsequently, the analytes were separated on a C18 UPLC column and detected by a tandem mass spectrometer. The method was validated using FDA guidelines. Finally, the validated assay was applied to the measurement of the plasma concentrations of 2q in plasma samples taken from mice after single intravenous doses of 2 mg/kg of 2q, and the pharmacokinetic parameters of 2q were determined.
Results
The calibration standards were linear (r2 ≥ 0.99) in the range of 1.56 – 200 nM of 2q with intra- and inter-run accuracy and precision values within the FDA guidelines. The lower limit of quantitation of the assay was 1.56 nM (0.258 pg on the column). The recoveries of 2q and IS from plasma were > 94%, with no appreciable matrix effect. The assay showed no significant carryover, and the plasma samples stored at –80oC or the processed samples stored in the autosampler at 10oC were stable for at least three weeks and 36 h, respectively. After intravenous injection, 2q showed a bi-exponential decline pattern in the mouse plasma, with a clearance of 30 mL/min/kg, a terminal volume of distribution of 1.93 mL/kg, and a terminal half-life of 45 min.
Conclusions
The developed assay is suitable for preclinical pharmacokinetic-pharmacodynamic studies of 2q as a potential drug candidate for ataxias.
Recommended Citation
Rahman, MA, Chandrashekar, DV, Nam, Y-W, et al. Development and validation of a UPLC-MS/MS method to investigate the plasma pharmacokinetics of a KCa2.2/KCa2.3 positive allosteric modulator in mice. Rapid Commun Mass Spectrom. 2023,37(15):e9537. https://doi.org/10.1002/rcm.9537
Copyright
Wiley
Included in
Medicinal and Pharmaceutical Chemistry Commons, Medicinal-Pharmaceutical Chemistry Commons
Comments
This is the accepted version of the following article:
Rahman, MA, Chandrashekar, DV, Nam, Y-W, et al. Development and validation of a UPLC-MS/MS method to investigate the plasma pharmacokinetics of a KCa2.2/KCa2.3 positive allosteric modulator in mice. Rapid Commun Mass Spectrom. 2023,37(15):e9537. https://doi.org/10.1002/rcm.9537
which will be published in final form at https://doi.org/10.1002/rcm.9537. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.