Ultrafast and Predictive Mass Spectrometry-Based Autotaxin Assays for Label-Free Potency Screening.
SLAS Discov
; 22(4): 425-432, 2017 Apr.
Article
em En
| MEDLINE
| ID: mdl-28328321
ABSTRACT
Autotaxin (ATX) is a promising drug target for the treatment of several diseases, such as cancer and fibrosis. ATX hydrolyzes lysophosphatidyl choline (LPC) into bioactive lysophosphatidic acid (LPA). The potency of ATX inhibitors can be readily determined by using fluorescence-based LPC derivatives. While such assays are ultra-high throughput, they are prone to false positives compared to assays based on natural LPC. Here we report the development of ultrafast mass spectrometry-based ATX assays enabling the measurement of data points within 13 s, which is 10 times faster than classic liquid chromatography-mass spectrometry. To this end, we set up a novel in vitro and whole-blood assay. We demonstrate that the potencies determined with these assays are in good agreement with the in vivo efficacy and that the whole-blood assay has the best predictive power. This high-throughput label-free approach paired with the translatable data quality is highly attractive for appropriate guidance of medicinal chemists for constructing strong structure-activity relationships.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Espectrometria de Massas
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Lisofosfolipídeos
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Lisofosfatidilcolinas
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Diester Fosfórico Hidrolases
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Inibidores Enzimáticos
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Ensaios de Triagem em Larga Escala
Tipo de estudo:
Diagnostic_studies
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Guideline
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Prognostic_studies
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Risk_factors_studies
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Screening_studies
Limite:
Animals
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Humans
Idioma:
En
Ano de publicação:
2017
Tipo de documento:
Article