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

Gordon, Laurie J.; Allen, Morven; Artursson, Per; Hann, Michael M.; Leavens, Bill; Mateus, André; Readshaw, Simon A.; Valkó, Klára; Wayne, Gareth; West, Andrew

doi:10.1177/1087057115604141

Cited by 57 publications

(56 citation statements)

References 13 publications

(23 reference statements)

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“…It was found that MB IAM values above 50 indicate promiscuous binding [57], and higher phospholipidotic potential [56][57][58][59] for discovery compound. It was also found that MB IAM showed a good correlation with the total cellular concentration of discovery compounds [60].…”

Section: Introductionmentioning

confidence: 86%

“…Recently, it was found that promiscuity (i.e., a discovery compound binding to multiple receptors) showed good correlation to the IAM MB of drug discovery compounds. The IAM MB data showed a good correlation with the intracellular concentration of compounds, indicating that membrane binding is important to get compounds into the cell [48]. Estimating the potential clinical dose as early as possible is an important aspect of the drug discovery process to help select the optimal compound profile and highest probability of successful progression to the clinic [49][50].…”

Section: Introductionmentioning

confidence: 99%

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In vitro membrane binding and protein binding (IAM MB/PB technology) to estimate in vivo distribution: applications in early drug discovery

Valkó¹,

Teague²,

Pidgeon³

2017

ADMET DMPK

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Section: Introductionmentioning

confidence: 86%

Section: Introductionmentioning

confidence: 99%

In vitro membrane binding and protein binding (IAM MB/PB technology) to estimate in vivo distribution: applications in early drug discovery

Valkó¹,

Teague²,

Pidgeon³

2017

ADMET DMPK

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“…First, it relies on sensitive analytical MS for compound quantification, which is considered a low-throughput technology. However, we recently showed that extensions of our methodology can process >1,000 compounds per week (29,30), which should be sufficient for secondary screening of hits from biochemical assays. Second, for compounds that need to be activated in the cell (prodrugs), the F ic of the dosed parent molecule might not be relevant.…”

Section: Discussionmentioning

confidence: 99%

“…Tissue-based methodologies allow accurate determinations of intracellular drug concentrations using unlabeled compounds, but they are based on animal tissues and have low throughput (26,27). Therefore, we developed a cell-based methodology for determination of intracellular drug concentrations in a high-throughput format, which is applicable to a wide variety of cell systems, not only those directly relevant for the target pharmacology (28)(29)(30). Our method does not require chemical labeling, has a high sensitivity, and measures the unbound drug concentration.…”

mentioning

confidence: 99%

Prediction of intracellular exposure bridges the gap between target- and cell-based drug discovery

Mateus

Gordon

Wayne

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Inadequate target exposure is a major cause of high attrition in drug discovery. Here, we show that a label-free method for quantifying the intracellular bioavailability (F ic ) of drug molecules predicts drug access to intracellular targets and hence, pharmacological effect. We determined F ic in multiple cellular assays and cell types representing different targets from a number of therapeutic areas, including cancer, inflammation, and dementia. Both cytosolic targets and targets localized in subcellular compartments were investigated. F ic gives insights on membrane-permeable compounds in terms of cellular potency and intracellular target engagement, compared with biochemical potency measurements alone. Knowledge of the amount of drug that is locally available to bind intracellular targets provides a powerful tool for compound selection in early drug discovery. intracellular drug bioavailability | drug exposure | target engagement | published kinase inhibitor set | MAPK14

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“…The coupling of a nano-LC system inline with a microfluidic chip to identify inhibitors of thrombin and factor Xa within complex mixtures of snake venom 8 623454J BXXXX10.1177/1087057115623454Journal of Biomolecular ScreeningWingfield and Wilson (2) demonstrates the versatility of MS and how it can be integrated with other assay platforms to deliver high-value data.…”

Section: From the Guest Editorsmentioning

confidence: 98%

Advances in Mass Spectrometry Within Drug Discovery

Wingfield

Wilson

2016

SLAS Discovery

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Mass spectrometry has evolved from a specialist technique, used initially almost exclusively for structure determination, to one where the bulk of applications now revolve around routine applications in analyte detection and quantification.In the beginning, samples of pure compounds were introduced into the ion source of the mass spectrometer (MS) deposited on a probe. Complex mixture analysis using probes was impractical, but could be performed by interfacing the MS with a gas chromatograph (GC). This was an early and highly successful development that enabled the analysis of compounds via first packed column, and then capillary GC-MS, but the technology was, obviously, limited to volatile compounds (or those that could be rendered volatile by derivatization). This requirement for thermally stable volatile analytes represented a clear limitation when the need was for the analysis of biological samples containing thermally labile, involatile analytes present in aqueous samples.From the point of view of those who struggled with the primitive, ineffective, and unreliable methods for interfacing MS with liquid chromatography (LC), such as the "moving belt" or direct liquid introduction methods, it sometimes seemed that the goal of producing a viable LC-MS system was going to be a step too far. However, this was before the development of the now dominant electrospray and atmospheric pressure chemical ionization (ESI and APCI) methods. These ionization techniques completely transformed the prospects for the analysis of involatile analytes in aqueous solution, which could now be performed using either direct liquid introduction (DLI) MS or LC-MS.In parallel to the hyphenation of LC with MS developments in surface analysis, techniques such as matrixassisted laser desorption ionization (MALDI) and desorption electrospray ionization (DESI) have produced tremendous opportunities for the direct analysis of samples deposited on a suitable support or for the high-resolution imaging of tissues.These advances in the ionization techniques available for MS combined with other developments in instrumentation, such as the introduction of triple-quadrupole, time-of-flight, and high-mass-resolution instruments (e.g., Fourier transform ion cyclotron resonance [FTICR] and "orbitraps"), as well as ion mobility, have enabled a suite of MS-based solutions to address complex problems within biology. It is not hard to see why MS-based methods have become popular, because when carefully optimized, mass spectrometry offers highly sensitive qualitative and quantitative analysis and specific detection combined with speed. Indeed, such has been the success of MS-based methodology that it is now difficult to envisage how modern bioscience, and particularly drug discovery and development, could be so efficiently conducted in its absence.Continual improvements in both the speed and sensitivity of MS-based techniques are facilitating an ever-increasing number of applications for high-throughput screening, and this is reflected in the content of thi...

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

Cited by 57 publications

References 13 publications

In vitro membrane binding and protein binding (IAM MB/PB technology) to estimate in vivo distribution: applications in early drug discovery

In vitro membrane binding and protein binding (IAM MB/PB technology) to estimate in vivo distribution: applications in early drug discovery

Prediction of intracellular exposure bridges the gap between target- and cell-based drug discovery

Advances in Mass Spectrometry Within Drug Discovery

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