Development of an UPLC–MS/MS method for quantification of Avitinib (AC0010) and its five metabolites in human cerebrospinal fluid: Application to a study of the blood-brain barrier penetration rate of non-small cell lung cancer patients

摘要

Highlights ? Avitinib (AC0010) is a novel mutant-selective EGFR-TKI that shows a good curative effect on NSCLC patients. ? Up to now, there has been no publication of determination method for Avitinib. ? Cerebrospinal fluid is a special matrix for which method development is difficult. ? The method determined not only the parent Avitinib, but also its five metabolites at the same time. ? The method successfully applied to the BBB penetration rate research of Avitinib for advanced NSCLC patients with metastatic encephaloma. Abstract Avitinib (AC0010) is a mutant-selective epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKI), designed to be a targeted therapeutic agent for non-small cell lung cancer (NSCLC) patients harboring EGFR active and T790M resistant mutations. A rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC–MS/MS) method was developed and validated for the determination of Avitinib and its five metabolites (M1, M2, M4, M7, MII-6) in human cerebrospinal fluid (CSF). The samples were purified by protein precipitation and separated on a BEH C 18 column (2.1 × 50 mm, 1.7 μm). Electrospray ionization (ESI) in positive ion mode and multiple reaction monitoring (MRM) were used to monitor the ion transitions at m / z 488/257, 474/403, 504/487, 434/377, 490/405, 476/391. The results indicated that the method had excellent sensitivity and specificity. The linear range covered from 0.05 to 50 ng/mL for Avitinib, M1, M4, M7, and MII-6, and from 0.01 to 10 ng/mL for M2. Intra-day and inter-day precisions (in terms of% RSD) were all 15% and the accuracies (in terms of% RE) were within the range of ±15%. The lower limit of quantification (LLOQ), matrix effect, extraction recovery, stability and dilution integrity were also validated and satisfied with the criteria of validation. Finally, the method was successfully applied to a blood-brain barrier (BBB) penetration rate research of NSCLC patients after an oral administration of Avitinib.