LC-HRMS and NMR studies for characterization of forced degradation impurities of ponatinib, a tyrosine kinase inhibitor, insights into in-silico degradation and toxicity profiles

• Published in: Journal of pharmaceutical and biomedical analysis 2023-04, Vol.227, p.115280-115280, Article 115280
• Main Authors: Golla, Vijaya Madhyanapu, Kushwah, Bhoopendra Singh, Dhiman, Vivek, Velip, Laximan, Samanthula, Gananadhamu
• Format: Article
• Language: English
• Subjects: Chromatography, High Pressure Liquid - methods; Degradation chemistry; Drug Stability; Forced degradation studies; in-silico toxicity studies; Magnetic Resonance Spectroscopy - methods; Mass spectrometry; Mass Spectrometry - methods; Nuclear magnetic resonance spectroscopy; Ponatinib; Tyrosine Kinase Inhibitors
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2023.115280

The degradation profile of ponatinib was established during the present study by exposing it to various stress conditions. In-silico degradation pattern of ponatinib was outlined by using Zeneth software. Five degradation impurities were formed during the stress testing of ponatinib. High performance liquid chromatographic method was developed to separate these degradation impurities which includes ammonium acetate of pH 4.75 (A) and methanol (B) as mobile phase in gradient elution mode and Waters Reliant C18 (4.6 × 250 mm, 5 µm) column as stationary phase. Optimised flow rate, injection volume and detection wavelength of the HPLC method were 1.0 mL/min, 10 µL and 254 nm, respectively. Chemical structures of degradation impurities were proposed by high resolution mass spectrometry further, major degradation products were isolated, enriched and investigated thoroughly with the aid of nuclear magnetic resonance spectroscopy studies. The degradation impurities were identified as 4-aminophthalaldehyde (DP 1), 4-((4-methylpiperazin-1-yl)methyl)− 3-(trifluoromethyl) benzenamine (DP 2), 3-(2-(imidazo[1,2-b]pyridazin-3-yl)acetyl)− 4-methylbenzoic acid (DP 3), 3-(2-(imidazo[1,2-b]pyridazin-3-yl)ethynyl)− 4-methylbenzoic acid (DP 4) and N-oxide impurity (DP 5) which are new and were not reported in the literature till date. Additionally, toxicity and mutagenicity profiles of ponatinib and its degradation impurities were predicted in-silico by using DEREK and SARAH software. This whole study gives meaningful insights about chemical stability of ponatinib which is useful in its drug development lifecycle. [Display omitted] •Forced degradation studies of ponatinib were performed as per ICH requirements.•A total of five degradation products were formed and are selectively separated by optimised HPLC method.•Five DPs formed were characterized by LC-HRMS.•Extensive NMR studies were executed on drug, DP 4 and DP 5.•In-silico degradation as well as toxicity profiles of ponatinib and its DPs were established.