Characterization of Alpelisib in Rat Plasma by a Newly Developed UPLC-MS/MS Method: Application to a Drug-Drug Interaction Study

Abstract

Alpelisib, an oral selective and small-molecule phosphoinositide 3-kinase inhibitor, was lately approved in the United States to treat breast cancer. A sensitive method to quantify alpelisib levels in rat plasma on the basis of ultra-performance liquid chromatography–tandem mass spectrometry technique was established and validated, which was successfully employed to explore the effects of CYP3A4 inhibitors on alpelisib pharmacokinetics in rats. A C18 column named Acquity UPLC BEH C18 was applied to achieve the separation of alpelisib and internal standard duvelisib after protein precipitation with acetonitrile. The mobile phase in this study had two components, namely, acetonitrile and water having 0.1% formic acid, and a program with gradient elution method was used at a flow rate of 0.40 ml/min. Mass spectrometry in a positive multiple reaction monitoring mode was operated. In the scope of 1–5,000 ng/ml, this assay had excellent linearity. Our newly developed assay was verified in all aspects of bioanalytical method validation, involving lower limit of quantification, selectivity, accuracy and precision, calibration curve, extraction recovery, matrix effect, and stability. Then, this assay was used to detect the plasma levels of alpelisib from a drug-drug interaction investigation, where ketoconazole remarkably increased the plasma concentration of alpelisib and changed alpelisib pharmacokinetics more than itraconazole. This study will help better understand the pharmacokinetic properties of alpelisib, and further clinical studies should be done to confirm this result in patients.

Bioanalysis of alpelisib using liquid chromatography–tandem mass spectrometry and application to pharmacokinetic study

“…To the best of our knowledge, this is the first study to report a method for overcoming the carryover effects in chromatographic analysis. In addition, improved symmetricity and reduced tailing effect were observed with this method compared to those in a previously reported study (Wang et al 2021).…”

“…Thus, the use of valid analytical methods to determine drug concentrations is inevitable in preclinical and clinical studies. To the best of our knowledge, valid analytical methods for the quantification of alpelisib in biological matrices have only been reported for rat plasma (Seo et al 2021;Wang et al 2021). However, large plasma sample volumes (100 µL) were used in these studies; therefore, the methods are not feasible for pharmacokinetic studies in mice, the most commonly used species for xenograft studies, considering that a small volume of blood is available from mice.…”

“…However, large plasma sample volumes (100 µL) were used in these studies; therefore, the methods are not feasible for pharmacokinetic studies in mice, the most commonly used species for xenograft studies, considering that a small volume of blood is available from mice. Although Wang et al (2021) applied a simplified pretreatment method for rat plasma compared to that reported by Seo et al (2021), improvement of the peak shape for alpelisib in a chromatogram is required owing to the shoulder peak to produce reproducible results. Several clinical studies on alpelisib have been reported (Mayer et al 2017;Juric et al 2018); however, to the best of our knowledge, there are no valid analytical methods for the quantification of alpelisib in human plasma.…”

Evaluation of the inhibitory effect of azoles on pharmacokinetics of lenvatinib in rats both in vivo and in vitro by UPLC‐MS/MS

“…Liu et al evaluated the drug interaction between isavuconazole and flumatinib by three sets of hepatic microsomal models, HLMs, RLMs and recombinant human CYP3A4, respectively 34 . It has been shown that both ketoconazole and itraconazole inhibit the in vivo metabolism of alpelisib 35 . Posazonazole differs from other triazole antifungals in that it is barely metabolized by the CYP450 pathway and 17% is glucuronidated by UGT1A4.…”

“…34 It has been shown that both ketoconazole and itraconazole inhibit the in vivo metabolism of alpelisib. 35 Posazonazole differs from other triazole antifungals in that it is barely metabolized by the CYP450 pathway and 17% is glucuronidated by UGT1A4. The inhibitory potency of posaconazole is reflected in the concentration.…”