Direct Measurement of Intracellular Compound Concentration by RapidFire Mass Spectrometry Offers Insights into Cell Permeability

Journal of Biomolecular Screening
Laurie J GordonAndy West

Abstract

One of the key challenges facing early stage drug discovery is understanding the commonly observed difference between the activity of compounds in biochemical assays and cellular assays. Traditionally, indirect or estimated cell permeability measurements such as estimations from logP or artificial membrane permeability are used to explain the differences. The missing link is a direct measurement of intracellular compound concentration in whole cells. This can, in some circumstances, be estimated from the cellular activity, but this may also be problematic if cellular activity is weak or absent. Advances in sensitivity and throughput of analytical techniques have enabled us to develop a high-throughput assay for the measurement of intracellular compound concentration for routine use to support lead optimization. The assay uses a RapidFire-MS based readout of compound concentration in HeLa cells following incubation of cells with test compound. The initial assay validation was performed by ultra-high performance liquid chromatography tandem mass spectrometry, and the assay was subsequently transferred to RapidFire tandem mass spectrometry. Further miniaturization and optimization were performed to streamline the process, increase...Continue Reading

References

Sep 11, 2009·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Gustav AhlinPer Artursson
Feb 8, 2011·Journal of Biomolecular Screening·Maureen K HighkinMarek M Nagiec
Jun 1, 2012·Expert Opinion on Drug Discovery·Dino MontanariTino Rossi
Aug 30, 2012·Journal of Pharmaceutical Sciences·Klara ValkoDino Montanari
May 2, 2013·Molecular Pharmaceutics·André MateusPer Artursson
Mar 8, 2014·Journal of Medicinal Chemistry·André MateusPer Artursson
Jun 3, 2014·Methods : a Companion to Methods in Enzymology·Michael M Hann, Graham L Simpson

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Citations

Oct 16, 2016·Bioprocess and Biosystems Engineering·Philippe JacquesRénato Froidevaux
Feb 22, 2017·PLoS Neglected Tropical Diseases·Antonio C B BurtolosoJosmar R Rocha
Jan 17, 2016·Journal of Biomolecular Screening·Jonathan Wingfield, Ian D Wilson
Feb 23, 2017·Scientific Reports·André MateusPer Artursson
Feb 12, 2017·Angewandte Chemie·Eric ValeurAlleyn T Plowright
May 10, 2018·Angewandte Chemie·Leila Peraro, Joshua A Kritzer
Jul 31, 2018·Clinical Pharmacology and Therapeutics·Yingying GuoUNKNOWN International Transporter Consortium
Sep 25, 2017·Nature Reviews. Drug Discovery·Andrew M DavisEric Valeur
Jul 14, 2017·Proceedings of the National Academy of Sciences of the United States of America·André MateusPer Artursson
Feb 2, 2019·Journal of Cheminformatics·Peter W Kenny
Dec 18, 2020·International Journal of Toxicology·Doris ZaneJessica Hawes
Sep 18, 2020·ACS Medicinal Chemistry Letters·Victoria G KleinR Scott Lokey

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Methods Mentioned

BETA
reverse-phase chromatography

Software Mentioned

Excel
DiscoveryQuant
RapidFire Integrator
Covaris
Analyst
RapidFire
DiscoverQuant Optimize and Analyze

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