An aqueous normal-phase chromatography coupled with tandem mass spectrometry method for determining unbound brain-to-plasma concentration ratio of AZD1775, a Wee1 kinase inhibitor, in patients with glioblastoma.
A rapid, sensitive, and robust aqueous normal-phase chromatography method coupled with tandem mass spectrometry was developed and validated for the quantitation of AZD1775, a Wee-1 inhibitor, in human plasma and brain tumor tissue. Sample preparation involved simple protein precipitation with acetonitrile. Chromatographic separation was achieved on ethylene bridged hybrid stationary phases (i.e., Waters XBridge Amide column) under an isocratic elution with the mobile phase consisting of acetonitrile/ammonium formate in water (10mM, pH 3.0) (85:15,v/v) at a flow rate of 0.8mL/min for 5min. The lower limit of quantitation (LLOQ) was 0.2ng/mL of AZD1775 in plasma and tissue homogenate. The calibration curve was linear over AZD1775 concentration range of 0.2-1000ng/mL in plasma and tissue homogenate. The intra- and inter-day precision and accuracy were within the generally accepted criteria for bioanalytical method (<15%). The method was successfully applied to assess the penetration of AZD1775 across the blood-brain tumor barrier, as assessed by the unbound brain-to-plasma ratio, in patients with glioblastoma.
Karmanos Cancer Institute, Wayne State University, Detroit, MI, 48201, USA.
Jianmei
J
Wu
Barrow Neurological Institute, St. Joseph's Hospital & Medical Center, Phoenix, AZ, 85013, USA.
Nader
N
Sanai
Karmanos Cancer Institute, Wayne State University, Detroit, MI, 48201, USA.
Xun
X
Bao
Yale Cancer Center, Yale School of Medicine, New Haven, CT, 06520, USA.
Patricia
P
LoRusso
Karmanos Cancer Institute, Wayne State University, Detroit, MI, 48201, USA. Electronic address: lijin@karmanos.org.
Jing
J
Li
D001921:Brain / Q000378:metabolism; D001932:Brain Neoplasms / Q000097:blood / Q000188:drug therapy / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005909:Glioblastoma / Q000097:blood / Q000188:drug therapy / Q000378:metabolism; D006801:Humans; D057230:Limit of Detection; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000097:blood / Q000378:metabolism / Q000493:pharmacokinetics*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011720:Pyrazoles / Q000097:blood / Q000378:metabolism / Q000493:pharmacokinetics*; D011743:Pyrimidines / Q000097:blood / Q000378:metabolism / Q000493:pharmacokinetics*; D011744:Pyrimidinones; D053719:Tandem Mass Spectrometry / Q000379:methods
D018797:Cell Cycle Proteins; D009687:Nuclear Proteins; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; D011743:Pyrimidines; D011744:Pyrimidinones; D011505:Protein-Tyrosine Kinases; C499349:WEE1 protein, human; C549567:adavosertib
AZD1775; Aqueous normal-phase chromatography; Fraction unbound; LC–MS/MS; Tandem mass spectrometry; Unbound brain-to-plasma ratio