Multiplexed LC-MS/MS method for the simultaneous quantitation of three novel hepatitis C antivirals, daclatasvir, asunaprevir, and beclabuvir in human plasma.

Abstract

Dual or triple combination regimens of novel hepatitis C direct-acting antivirals (DAA, daclatasvir, asunaprevir, or beclabuvir) provide high sustained virological response rates and reduced frequency of resistance compared to clinical monotherapy. To support pharmacokinetic (PK) assessments in clinical studies, a multiplexed liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous quantitation of daclatasvir, asunaprevir, beclabuvir (BMS-791325) and its active metabolite (BMS-794712) in human plasma was developed and validated. Human plasma samples were extracted with methyl-t-butyl ether followed by an LC-MS/MS analysis, which was conducted in a multiple reaction monitoring (MRM) mode. The lower limits of quantitation (LLOQ) were 1 ng/mL for daclatasvir, asunaprevir, and BMS-794712, and 2 ng/mL for beclabuvir. Intra-run precision (≤4.5% CV), inter-run precision (≤2.9% CV), and accuracy (±5.3% deviation) based on different concentration levels (low, geometric mean, mid and high) of the quality control samples (QCs) provided evidence of the methods accuracy and precision. Selectivity and matrix effect on LC-MS/MS detection, stability in plasma, and potential interference of coadministered drugs (ribavirin and interferon) were all evaluated and the results were acceptable. Method reproducibility was demonstrated by the reanalysis of a portion of study samples. The cross-validation results for QCs demonstrated the equivalency between this method and two single-analyte methods which were previously validated for quantitation of daclatasvir in human plasma. This approach of using a multiplexed LC-MS/MS method for the simultaneous quantitation of three DAAs is time- and cost-effective, and can maintain good data quality in sample analysis.

• Authors: Hao Jiang, Hamza Kandoussi, Jianing Zeng, Jian Wang, Roger Demers, Timothy Eley, Bing He, Richard Burrell, John Easter, Pathanjali Kadiyala, Janice Pursley, Laura Cojocaru, Chanda Baker, John Ryan, Anne-Françoise Aubry, Mark E Arnold
• Journal: Journal of pharmaceutical and biomedical analysis (J Pharm Biomed Anal) Vol. 107 Pg. 409-18 (Mar 25 2015) ISSN: 1873-264X [Electronic] England
• PMID: 25676854 (Publication Type: Journal Article)
• Copyright: Copyright © 2015 Elsevier B.V. All rights reserved.

Chemical References

• 8-cyclohexyl-N-((dimethylamino)sulfonyl)-1,1a,2,12b-tetrahydro-11-methoxy-1a-((3-methyl-3,8-diazabicyclo(3.2.1)oct-8-yl)carbonyl)cycloprop(d)indolo(2,1-a)(2)benzazepine-5-carboxamide
• Antiviral Agents
• Benzazepines
• Carbamates
• Imidazoles
• Indoles
• Isoquinolines
• Pyrrolidines
• Sulfonamides
• Ribavirin
• Interferons
• Valine
• daclatasvir
• asunaprevir

Topics

• Antiviral Agents (blood, chemistry, pharmacology)
• Benzazepines (blood, chemistry)
• Carbamates
• Chromatography, Liquid (methods)
• Hepacivirus (drug effects)
• Humans
• Imidazoles (blood, chemistry)
• Indoles (blood, chemistry)
• Interferons (blood, chemistry)
• Isoquinolines (blood, chemistry)
• Plasma (chemistry)
• Pyrrolidines
• Reproducibility of Results
• Ribavirin (blood, chemistry)
• Sulfonamides (blood, chemistry)
• Tandem Mass Spectrometry (methods)
• Valine (analogs & derivatives)