Characterization of forced degradation products of pazopanib hydrochloride by UHPLC‐Q‐TOF/MS and in silico toxicity prediction

Abstract

Pazopanib (PZ), an anti-cancer drug, was subjected to forced degradation under hydrolytic (acid, base and neutral), oxidative, photolytic and thermal stress conditions as per International Conference on Harmonization guidelines. A selective stability indicating validated method was developed using a Waters Acquity UPLC HSS T3 (100 × 2.1 mm, 1.7 µm) column in gradient mode with ammonium acetate buffer (10 mM, pH 5.0) and acetonitrile. PZ was found to degrade only in photolytic conditions to produce six transformation products (TPs). All the TPs were identified and characterized by liquid chromatography/atmospheric pressure chemical ionization-quadrupole-time of flight mass spectrometry experiments in combination with accurate mass measurements. Plausible mechanisms have been proposed for the formation of TPs. In silico toxicity was predicted using TOPKAT and DEREK softwares for all the TPs. The TP, N4-(2,3-dimethyl-2H-indazol-6-yl)-N4-methylpyrimidine-2,4-diamine, was found to be genotoxic, whereas all other TPs with sulfonamide moiety were hepatotoxic. The data reported here are expected to be of significance as this study foresees the formation of one potential genotoxic and five hepatotoxic degradation/transformation products.

Metalloporphyrin-Catalyzed Oxidation of Sunitinib and Pazopanib, Two Anticancer Tyrosine Kinase Inhibitors: Evidence for New Potentially Toxic Metabolites

“…In vitro, the drug was found stable under very drastic conditions: 3 M HCl, 3 M NaOH at 80 °C or 30% H 2 O 2 at room temperature for 2 days. 20 Pazopanib was susceptible only to photolytic conditions (UV light for 3 days at 200 W•h/m 2 ). 20 In these conditions, the oxidation of H 3 C−C7 to an aldehyde function was proposed on the basis of UHPLC-MS fragmentation patterns.…”

“…20 Pazopanib was susceptible only to photolytic conditions (UV light for 3 days at 200 W•h/m 2 ). 20 In these conditions, the oxidation of H 3 C−C7 to an aldehyde function was proposed on the basis of UHPLC-MS fragmentation patterns. However, these studies, carried out very far from any biologically relevant conditions, do not allow to propose relevant in vivo metabolism of this drug.…”

“…After administration of [ 14 C]­pazopanib to 10 patients, on the basis of HPLC-MS analyses, hydroxylations of the drug at C7 and C19 positions, along with N8-demethylation, have been proposed as P450-mediated phase I metabolism. In vitro , the drug was found stable under very drastic conditions: 3 M HCl, 3 M NaOH at 80 °C or 30% H 2 O 2 at room temperature for 2 days . Pazopanib was susceptible only to photolytic conditions (UV light for 3 days at 200 W·h/m 2 ) .…”

A metabolomic perspective of pazopanib-induced acute hepatotoxicity in mice

“…The neutral cleavage of O -moiety suggested the formation of pazopanib N-oxide. The cleavage of formaldehyde, which was associated with a Meisenheimer rearrangement reaction (the methyl transfect from nitrogen atom into oxygen atom, N-CH 3 → O-CH 3 ) to generate the ion at m/z 424 + , indicating that the N -oxide reaction formed in the terminal nitrogen atom linked with the methyl group (Patel et al, 2015). The patterns of MS/MS fragmentation of M1 , M2 and M3 are provided in Figure 3(D–F).…”

Analytical Method Development and Validation of RP-HPLC Method for Estimation of Pazopanib Drug Sample and It’s Dosage Form

“…Studies have utilized sophisticated analytical methods, such as LC-MS/MS Verheijen et al, 2018;Minocha et al, 2012;Sparidans et al, 2012;Pressiat et al, 2018) UPLC-QTOF/MS (Patel et al, 2015), and High-Performance Liquid Chromatography -UV (Escudero-Ortiz et al, 2015;Sharada et al, 2016), to quantify pharmaceuticals. In addition, Ultra Violet techniques (Sharada and Babu, 2016;Chaitanya and Pawar, 2015) have been used to estimate this medication.…”