Characterization of forced degradation products of pazopanib hydrochloride by UHPLC-Q-TOF/MS and in silico toxicity prediction.
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.
Department of Pharmaceutical Analysis, National Institute of Pharmaceutical Education and Research, Balanagar, Hyderabad, 500037, Telangana, India.
Prinesh N
PN
Patel
Department of Pharmaceutical Analysis, National Institute of Pharmaceutical Education and Research, Balanagar, Hyderabad, 500037, Telangana, India.
Pradipbhai D
PD
Kalariya
Department of Pharmacoinformatics, National Institute of Pharmaceutical Education and Research, Sector 67, SAS Nagar (Mohali), Punjab, 160062, India.
Mahesh
M
Sharma
Department of Pharmacoinformatics, National Institute of Pharmaceutical Education and Research, Sector 67, SAS Nagar (Mohali), Punjab, 160062, India.
Prabha
P
Garg
Department of Pharmaceutical Analysis, National Institute of Pharmaceutical Education and Research, Balanagar, Hyderabad, 500037, Telangana, India.
M V N Kumar
MV
Talluri
Department of Pharmaceutical Analysis, National Institute of Pharmaceutical Education and Research, Balanagar, Hyderabad, 500037, Telangana, India.
S
S
Gananadhamu
Department of Pharmaceutical Analysis, National Institute of Pharmaceutical Education and Research, Balanagar, Hyderabad, 500037, Telangana, India.|National Center for Mass Spectrometry, CSIR-Indian Institute of Chemical Technology, Hyderabad, 500 007, Telangana, India.
R
R
Srinivas
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003198:Computer Simulation; D004355:Drug Stability; D005260:Female; D007191:Indazoles; D057230:Limit of Detection; D008297:Male; D051379:Mice; D009153:Mutagens / Q000737:chemistry* / Q000633:toxicity*; D011743:Pyrimidines / Q000737:chemistry* / Q000633:toxicity*; D051381:Rats; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D013449:Sulfonamides / Q000737:chemistry* / Q000633:toxicity*
D007191:Indazoles; D009153:Mutagens; D011743:Pyrimidines; D013449:Sulfonamides; C516667:pazopanib
LC-MS/MS; UHPLC stability assay; forced degradation; genotoxic transformation products; pazopanib; stress studies