Development and validation of a multiplex UHPLC-MS/MS method for the determination of the investigational antibiotic against multi-resistant tuberculosis macozinone (PBTZ169) and five active metabolites in human plasma

Autor: Vincent Desfontaine, Laurent A. Decosterd, Vadim Makarov, Anthony Vocat, O. B. Ryabova, Jeff Pitteloud, Stewart T. Cole, Anton Ivanyuk, Sylvie Guinchard, Sandra Cruchon, Dany Spaggiari, Thierry Buclin, Emilyne Blattes, Lorenzo Ciullini, Carine Bardinet
Jazyk: angličtina
Rok vydání: 2019
Předmět:
Male
Metabolic Processes
Physiology
Metabolite
Antitubercular Agents
Thiazines
sfstp proposal
Biochemistry
Piperazines
combination therapy
chemistry.chemical_compound
Spectrum Analysis Techniques
Drug Metabolism
Tandem Mass Spectrometry
Tuberculosis
Multidrug-Resistant
Medicine and Health Sciences
Metabolites
Multiplex
Chromatography
High Pressure Liquid
0303 health sciences
Multidisciplinary
biology
Chemistry
benzothiazinones
Anti-Bacterial Agents
Body Fluids
Actinobacteria
Blood
strategies
Metabolome
Medicine
Anatomy
Switzerland
Research Article
Bioanalysis
Analyte
bioanalysis
Science
Context (language use)
mycobacterium-tuberculosis
Research and Analysis Methods
Blood Plasma
Mycobacterium tuberculosis
03 medical and health sciences
Pharmacokinetics
Humans
Active metabolite
030304 developmental biology
Pharmacology
Chromatography
Bacteria
030306 microbiology
Organisms
Reproducibility of Results
Biology and Life Sciences
Inductively Coupled Plasma Emission Spectroscopy
quantitative analytical procedures
Drugs
Investigational
biology.organism_classification
Metabolism
harmonization
identification
chromatography-mass-spectrometry
Zdroj: PLoS ONE, Vol 14, Iss 5, p e0217139 (2019)
PLoS ONE
PloS one, vol. 14, no. 5, pp. e0217139
ISSN: 1932-6203
Popis: The emergence of Mycobacterium tuberculosis strains resistant to current first-line antibiotic regimens constitutes a major global health threat. New treatments against multidrug-resistant tuberculosis (MDR-TB) are thus eagerly needed in particular in countries with a high MDR-TB prevalence. In this context, macozinone (PBTZ169), a promising drug candidate with an unique mode of action and highly potent in vitro tuberculocidal properties against MDR Mycobacterium strains, has now reached the clinical phase and has been notably tested in healthy male volunteers in Switzerland. To that endeavor, a multiplex UHPLC-MS/MS method has been developed for the sensitive and accurate human plasma levels determination of PBTZ169 along with five metabolites retaining in vitro anti-TB activity. Plasma protein precipitation with methanol was carried out as a simplified sample clean-up procedure followed by direct injection of the undiluted supernatant for the bioanalysis of the six analytes within 5 min, using 1.8 mu m reversed-phase chromatography coupled to triple quadrupole mass spectrometry employing electrospray ionization in the positive mode. Stable isotopically-labelled PBTZ169 was used as internal standard (ISTD), while metabolites could be reliably quantified using two unlabeled chemical analogues selected as ISTD from a large in-house analogous compounds library. The overall methodology was fully validated according to current recommendations (FDA, EMEA) for bioanalytical methods, which include selectivity, carryover, qualitative and quantitative matrix effect, extraction recovery, process efficiency, trueness, precision, accuracy profiles, method and instrument detection limits, integrity to dilution, anticoagulant comparison and short-and long-term stabilities. Stability studies on the reduced metabolite H-2-PBTZ169 have shown no significant impact on the actual PBTZ169 concentrations determined with the proposed assay. This simplified, rapid, sensitive and robust methodology has been applied to the bioanalysis of human plasma samples collected within the frame of a phase I clinical study in healthy volunteers receiving PBTZ169.
Databáze: OpenAIRE
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