Quantitative analysis of D‐24851, a novel anticancer agent, in human plasma and urine by liquid chromatography coupled with tandem mass spectrometry

Abstract

The development of a liquid chromatography/tandem mass spectrometric assay for the quantitative analysis of the novel tubulin inhibitor D-24851 in human plasma and urine is described. D-24851 and the deuterated internal standard were extracted from 250 microL of plasma or urine using hexane/ether (1:1, v/v). Subsequently, 10-microL aliquots of reconstituted extracts were injected onto an Inertsil ODS analytical column (50 x 2.0 mm i.d., 5 microm particle size). An eluent consisting of methanol/5 mM ammonium acetate, 0.004% formic acid in water (80:20, v/v) was pumped at a flow rate of 0.2 mL/min. An API 365 triple quadrupole mass spectrometer was used in the multiple reaction monitoring mode for sensitive detection. For human plasma a dynamic range of 1-1000 ng/mL was validated, and for human urine a range of 0.25-50 ng/mL. Validation was performed according to the most recent FDA guidelines and all results were within requirements. The assay has been successfully applied to support a phase I clinical trial with orally administered D-24851.

Stable isotopically labeled internal standards in quantitative bioanalysis using liquid chromatography/mass spectrometry: necessity or not?

“…3) only an analogous internal standard was available, lacking the chlorine atom at the benzyl moiety. 8 This internal standard appeared inappropriate for use in the assay, as is demonstrated in Table 1 (accuracy >15%, precision >15%). Quantitation without the use of an internal standard yielded even better results.…”

“…The quadruply deuterated SIL internal standard yielded excellent results (Table 1). 8 For the LC/ESI-MS/MS assay of the depsipeptide marine anticancer agent Kahalalide F, which contains a 5-methylhexanoic acid conjugated to the N-terminus, a butyric acid analogue was available as internal standard (Fig. 4).…”

Phase I dose-finding and pharmacokinetic trial of orally administered indibulin (D-24851) to patients with solid tumors

“…Indibulin induces accumulation of cells with condensed nuclei and abnormal mitotic spindles and arrests cells at the metaphase. The drug binds to tubulin and competes with colchicine, podophyllotoxin and nocodazole, but not with paclitaxel or vinblastine [2][3][4]. Furthermore, in vitro studies showed that indibulin blocks migration and inhibits growth of endothelial cells, suggesting antiangiogenic properties [3].…”

Dose-finding and pharmacokinetic study of orally administered indibulin (D-24851) to patients with advanced solid tumors

“…Indibulin was stable at −20°C in plasma over the time-period stored and analyzed, and not influenced by freeze-thawing [12]. Indibulin plasma concentrations were measured by a validated LC-MS/MS method [12]. The lower limit of quantitation was 1.00 ng/ mL for plasma.…”

“…Samples were centrifuged at 3,000 g for 10 min at room temperature and plasma was stored at −20°C until analysis. Indibulin was stable at −20°C in plasma over the time-period stored and analyzed, and not influenced by freeze-thawing [12]. Indibulin plasma concentrations were measured by a validated LC-MS/MS method [12].…”