Analytical Evaluation of MCC950 Brain Penetration in a Rodent Model using LC-MS/MS

Abstract

MCC950, a selective NLRP3 inflammasome inhibitor, holds significant potential for the treatment of neuroinflammatory diseases. This study focuses on developing a robust analytical method to evaluate the ability of MCC950 to cross the blood-brain barrier (BBB) and accumulate in brain tissue after administration to rodents. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method using negative mode electrospray ionization (ESI) was developed to accurately and rapidly detect MCC950. The analysis was performed on a ThermoFisher Vanquish UHPLC coupled to a TSQ Altis mass spectrometer using a C18 column and a gradient elution of 0.1% formic acid in deionized water and acetonitrile. Quantitative detection was achieved with an MRM transition of m/z 403 → 204, indicating the loss of sodium (Na？) from MCC950 sodium (exact mass: 426.122). Preprocessing included extraction of brain tissue samples with MTBE, centrifugation, and subsequent reconstitution in a solvent mixture of formic acid and acetonitrile. Control samples receiving only vehicle (PBS) showed no detectable MCC950, while the experimental samples confirmed the presence of the compound in brain tissue. These results demonstrate that MCC950 successfully crosses the BBB and that the LC-MS/MS method developed provides a reliable platform for assessing drug distribution in the central nervous system. This analytical approach improves our understanding of the pharmacokinetics of MCC950 and supports its further development as a therapeutic for neuroinflammation.