Liquid chromatography–tandem mass spectrometric assay for the quantitation of the novel radiation protective agent and radiation mitigator JP4-039 in murine plasma

Abstract

JP4-039 radio-protects prior to, and radio-mitigates after ionizing radiation by neutralizing reactive oxygen species. We developed and validated an LC-MS/MS assay for the quantitation of JP4-039 in murine plasma. Methanol protein precipitation of 50μL plasma was followed by isocratic reverse phase chromatography for a 6min run time, and electrospray positive mode ionization mass spectrometric detection. The plasma assay was linear from 1 to 1000ng/mL with appropriate accuracy (97.1-107.6%) and precision (3.7-12.5%CV), and fulfilled FDA guidance criteria. Recovery was 77.2-136.1% with moderate ionization enhancement (10.9-39.5%). Plasma freeze-thaw stability (98.8-104.2%), stability for 13.5 months at -80°C (93.1-105.6%), and stability for 4h at room temperature (94.2-97.6%) were all acceptable. Limited cross-validation to tissue homogenates suggested that these could also be analyzed for JP4-039 accurately. This assay has been directly applied to determine the pharmacokinetics of JP4-039 in C57BL/6 male mice after IV administration of 20mg/kg JP4-039 and will be extended to other studies of this agent.

The mitochondrial‐targeted reactive species scavenger JP4‐039 prevents sulfite‐induced alterations in antioxidant defenses, energy transfer, and cell death signaling in striatum of rats

“…Changes in MAPK and apoptosis markers were prevented or attenuated by JP4‐039, which is consistent with prior publications showing that ROS may modulate phosphorylation of MAPKs 49,50 and trigger mitochondrial permeability transition 51 . These findings are of particular interest given that JP4‐039 and closely related analogs, such as XJB‐5‐131, have been shown to cross the blood‐brain barrier in mice, and that both iSOD and MoCD are characterized by severe neurological dysfunction and basal ganglia abnormalities 47,52 …”

ETHE1 and MOCS1 deficiencies: Disruption of mitochondrial bioenergetics, dynamics, redox homeostasis and endoplasmic reticulum-mitochondria crosstalk in patient fibroblasts

“…In the current study, we also verify that JP4-039 ameliorates ROS accumulation in ETHE1 and MOCS1 deficient fibroblasts and improves mitochondrial respiration. These findings are particularly interesting since previous data showed that JP4-039 crosses brain-blood barrier in mice 78 and ETHE1 and MOCS1 deficiencies are inborn errors mainly characterized by neurological dysfunction. Furthermore, N -acetylcysteine, an antioxidant currently used to treat ETHE1 deficient patients 10,13,79,80 , did not affect superoxide levels, demonstrating the importance of mitochondrial targeting for the biological efficacy of free radical scavengers.…”

Evaluation of Different Formulations and Routes for the Delivery of the Ionizing Radiation Mitigator GS-Nitroxide (JP4-039)

“…effective in mitigating irradiation induced damage. The PK of achieving whole blood levels were significantly delayed compared to that seen with IV administration (27), although effective radiation mitigation was also achieved. The surface area required for effective administration of JP4-039 by MNA required exposure of a significant volume of skin.…”

“…Pharmacokinetic (PK) studies. The methods for liquid chromatography-tandem Mass Spectrophotometric analysis of mouse plasma for levels of JP4-039 have been previously described (27).…”

“…Previous studies have demonstrated safe delivery of doses of JP4-039 as low as 5 mg/kg and up to 20 mg/kg, delivered in Cremophor EL/EtOH. Minimum toxicity has been detected at the dose of 30 mg/kg (27). IV delivery of JP4-039 in Cremophor/ethanol was compared to the previously published formulation F14 (13,28,31).…”