A novel uHPLC-MS/MS method for the quantitation of AZD7451 (AZ12607092) in human plasma.
Tropomyosin-related kinases (Trk) are tyrosine kinase receptors implicated in tumor proliferation, invasion, and survival signaling across a number of tumors, making them potentially attractive targets for the treatment of cancer. AZD7451 is a potent and selective inhibitor of Trk kinases currently undergoing a Phase I dose escalation in glioblastoma multiforme at the National Cancer Institute. A key part of early clinical testing for AZD7451 involves demonstrating that pharmacokinetic half-life and clinical exposures of AZD7451 are sufficient to inhibit Trk receptors in preclinical models. To address this need, an ultra sensitive analytical method was developed to measure the AZD7451 profile in human plasma. A liquid-liquid extraction recovered >80% of AZD7451 before quantitative analysis by ultra HPLC-MS/MS. A Varian Polaris(®) C18-A column and a mass transition of m/z 383.5→340.5 (m/z 389.6→342.0 for the internal standard [(2)H6]-AZD7451) was used, and a dynamic calibration range of 0.5-1000ng/mL was established, which provided a sensitive (<8.5% deviation), and precise (<6%) quantitative assay for AZD7451. AZD7451 demonstrated stability in human plasma at room temperature for 24h (<7% change) and after extraction at 4°C for 24h (<8% change), and was stable through 4 freeze/thaw cycles (<8% change). This method was used to measure AZD7451 plasma levels in clinical samples to confirm the sensitivity at several time points following AZD7451 treatment in subjects with glioblastoma.
Clinical Pharmacology Program, Office of the Clinical Director, National Cancer Institute, Bethesda, MD, United States.
Cody J
CJ
Peer
Jeffrey L
JL
Brown
Timothy J
TJ
Martin
Jeffrey
J
Roth
Shawn D
SD
Spencer
Patrick
P
Brassil
Katharine A
KA
McNeill
Teri N
TN
Kreisl
Howard A
HA
Fine
William D
WD
Figg
D015075:2-Aminopurine / Q000097:blood* / Q000737:chemistry / Q000031:analogs & derivatives; D000970:Antineoplastic Agents / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D016014:Linear Models; D047428:Protein Kinase Inhibitors / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D011720:Pyrazoles / Q000097:blood* / Q000737:chemistry; D020794:Receptor Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
C000589263:AZD7451; D000970:Antineoplastic Agents; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; D015075:2-Aminopurine; D020794:Receptor Protein-Tyrosine Kinases
LLOQ; MS; MS/MS; QC; Tandem mass spectrometry; Tropomyosin-related kinase; Ultra-high performance liquid chromatography; lower limit of quantification; mass spectrometry; quality control; tandem mass spectrometry; uHPLC; ultra-high performance liquid chromatography