title: Validation of a liquid chromatography-triple quadrupole mass spectrometric method for the determination of 5-nitro-5'-hydroxy-indirubin-3'-oxime (AGM-130) in human plasma and its application to microdose clinical trial.
abstract: A liquid chromatography-triple quadrupole mass spectrometric (LC-MS/MS) method was developed and validated for the determination of 5-nitro-5'-hydroxy-indirubin-3'-oxime (AGM-130) in human plasma to support a microdose clinical trial. The method consisted of a liquid-liquid extraction for sample preparation and LC-MS/MS analysis in the positive ion mode using TurboIonSpray(TM) for analysis. d3 -AGM-130 was used as the internal standard. A linear regression (weighted 1/concentration) was used to fit calibration curves over the concentration range of 10-2000 pg/mL for AGM-130. There were no endogenous interference components in the blank human plasma tested. The accuracy at the lower limit of quantitation was 96.6% with a precision (coefficient of variation, CV) of 4.4%. For quality control samples at 30, 160 and 1600 pg/mL, the between run CV was ≤5.0 %. Between-run accuracy ranged from 98.1 to 101.0%. AGM-130 was stable in 50% acetonitrile for 168 h at 4°C and 6 h at room temperature. AGM-130 was also stable in human plasma at room temperature for 6 h and through three freeze-thaw cycles. The variability of selected samples for the incurred sample reanalysis was ≤12.7% when compared with the original sample concentrations. This validated LC-MS/MS method for determination of AGM-130 was used to support a phase 0 microdose clinical trial.
authors.affiliation: College of Pharmacy, Chungnam National University, Daejeon, 305-764, Republic of Korea.
authors.forename: Min-Ho
authors.initials: MH
authors.lastname: Park
authors.affiliation: College of Pharmacy, Chung-Ang University, Seoul, 156-756, Republic of Korea.|Bio-Core Ltd, Seoul, 153-760, Republic of Korea.
authors.forename: Yun Young
authors.initials: YY
authors.lastname: Lee
authors.affiliation: Bio-Core Ltd, Seoul, 153-760, Republic of Korea.
authors.forename: Kyung Hee
authors.initials: KH
authors.lastname: Cho
authors.affiliation: Bio-Core Ltd, Seoul, 153-760, Republic of Korea.
authors.forename: Sookie
authors.initials: S
authors.lastname: La
authors.affiliation: Bio-Core Ltd, Seoul, 153-760, Republic of Korea.
authors.forename: Hee Joo
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: College of Medicine, Catholic University of Korea, Seoul, 137-701, Republic of Korea.
authors.forename: Dong-Seok
authors.initials: DS
authors.lastname: Yim
authors.affiliation: School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju, 500-712, Republic of Korea.|Division of Drug Discovery, Anygen Co., Ltd, 500-712, Republic of Korea.
authors.forename: Sooho
authors.initials: S
authors.lastname: Ban
authors.affiliation: School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju, 500-712, Republic of Korea.|Division of Drug Discovery, Anygen Co., Ltd, 500-712, Republic of Korea.
authors.forename: Moon-Young
authors.initials: MY
authors.lastname: Park
authors.affiliation: School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju, 500-712, Republic of Korea.|Division of Drug Discovery, Anygen Co., Ltd, 500-712, Republic of Korea.
authors.forename: Yong-Chul
authors.initials: YC
authors.lastname: Kim
authors.affiliation: College of Pharmacy, Dankook University, Cheonan, 330-714, Republic of Korea.
authors.forename: Yoon-Gyoon
authors.initials: YG
authors.lastname: Kim
authors.affiliation: College of Pharmacy, Chungnam National University, Daejeon, 305-764, Republic of Korea.
authors.forename: Young G
authors.initials: YG
authors.lastname: Shin
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors; D005260:Female; D006801:Humans; D007211:Indoles / Q000008:administration & dosage* / Q000097:blood* / Q000493:pharmacokinetics; D016014:Linear Models; D059625:Liquid-Liquid Extraction; D008297:Male; D010091:Oximes / Q000008:administration & dosage* / Q000097:blood* / Q000493:pharmacokinetics; D047428:Protein Kinase Inhibitors / Q000008:administration & dosage / Q000097:blood / Q000493:pharmacokinetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: C000609730:5-nitro-5'-hydroxyindirubin-3'-oxime; D007211:Indoles; D010091:Oximes; D047428:Protein Kinase Inhibitors; D018844:Cyclin-Dependent Kinases
keywords: AGM-130; LC-MS/MS; cyclin-dependent kinase (CDK) inhibitor; liquid-liquid extraction; microdose clinical trial