title: High-performance liquid chromatography analysis of a novel small-molecule, anti-cancer drug, Palomid 529, in human and mouse plasma and in mouse tissue homogenates.
abstract: Palomid 529 (8-(1-Hydroxy-ethyl)-2-methoxy-3-(4-methoxy-benzyloxy)-benzo[c]chromen-6-one), is a novel non-steroidal small-molecule drug, which inhibits both mTORC1 and mTORC2 assembly, and elicits both anti-angiogenic and direct anti-tumor effects in vivo. We have developed and validated a sensitive and selective method for the quantification of Palomid 529 in human and mouse plasma and in a range of mouse tissue samples. Sample pretreatment involved liquid-liquid extraction with tert-butyl methyl ether yielding a recovery of >75%. Palomid 529 and the internal standard Palomid 545 were separated using a GraceSmart RP18 column (2.1 mm × 150 mm) packed with 5 μm C-18 material and a mobile phase comprised of 50% (v/v) acetonitrile and 50% (v/v) water delivered at a flow rate of 0.2 ml/min, and were detected by UV absorbance at a wavelength of 315 nm. Within the linear range of the calibration curve (10-10,000 ng/ml), acceptable accuracy and precision was achieved for all tested matrices. The validation results show that the method was selective and reproducible. Palomid 529 was stable in plasma upon 3 repeated freeze-thaw cycles and during storage for up to 24h at ambient temperature. However, pre-treated samples waiting for HPLC analyses need to be kept under dimmed light conditions at ambient temperature since a significant degradation of both Palomid 529 and Palomid 545 was observed when exposed to light. A pilot pharmacokinetic study in mice demonstrated the applicability of this method for pharmacokinetic purposes. Even at a low dose of 5.4 mg/kg this assay was still sensitive enough to determine the drug concentration in plasma samples obtained up to 24h after administration.
authors.affiliation: Department of Clinical Chemistry/Preclinical Pharmacology, The Netherlands Cancer Institute (Antoni van Leeuwenhoek Hospital), Amsterdam, The Netherlands.
authors.forename: Fan
authors.initials: F
authors.lastname: Lin
authors.forename: David
authors.initials: D
authors.lastname: Sherris
authors.forename: Jos H
authors.initials: JH
authors.lastname: Beijnen
authors.forename: Olaf
authors.initials: O
authors.lastname: Van Tellingen
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000032:analysis* / Q000097:blood / Q000737:chemistry; D001578:Benzopyrans / Q000032:analysis* / Q000097:blood / Q000737:chemistry; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D007668:Kidney / Q000737:chemistry; D016014:Linear Models; D059625:Liquid-Liquid Extraction; D008099:Liver / Q000737:chemistry; D008168:Lung / Q000737:chemistry; D051379:Mice; D009206:Myocardium / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013154:Spleen / Q000737:chemistry; D013696:Temperature
chemical_list: D000970:Antineoplastic Agents; D001578:Benzopyrans; C533469:palomid 529