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Miniaturization of two high throughput metabolic stability assays in early drug discovery

Miniaturization of two high throughput metabolic stability assays in early drug discovery

Beatrice D'Aubigné
Göteborg : Chalmers tekniska högskola, 2018. 44 s.
[Examensarbete på avancerad nivå]

When it was recognized that evaluation of pharmacokinetic (PK) properties during drug discovery strongly influenced the success rate during clinical development, ADME (Absorption, Distribution, Metabolism, Excretion) properties gained more and more attention in the pharmaceutical industry. One of the important PK properties to investigate is metabolic clearance since it directly relates to drug elimination and bioavailability. This master’s thesis project was performed at AstraZeneca whereinvitrometabolicstabilityisscreenedinearlydrugdevelopmentusinghuman liver microsomes and cryopreserved rat hepatocytes. The assays are run in 96-well format and the aim of this project was to miniaturize the two assays to 384-well format. Different equipment was tested when performing the experiments and the incubation volumes were optimized. The developed assay using human liver microsomes was validated in two steps. First, intra-assay and inter-assay variability was determinedandtheassaywascomparedwiththeexisting96-wellformatassay. The resulting data showed that AFE was 1.30 and AAFE 1.33. Then, the data was used to investigate the in vivo prediction and the results showed that the data align well with in vivo clearance data. The CLint values obtained from the developed assay using hepatocytes were on average lower than the CLint values obtained from the existing assay. Adding shaking decreased AFE from 2.19 to 1.29 and AAFE from 2.52 to 1.62 but seven of the 19 tested compounds did still have an average fold change ≥ 2. Lack of sufficient mixing during incubation was probably the main reason.

Nyckelord: Metabolic stability, CLint, human liver microsomes, rat hepatocytes, IVIVE

Publikationen registrerades 2018-12-17. Den ändrades senast 2018-12-17

CPL ID: 256409

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