Development, optimization and implementation of a centralized metabolic soft spot assay

Bioanalysis. 2017 Apr;9(7):541-552. doi: 10.4155/bio-2016-0299. Epub 2017 Mar 24.

Abstract

Aim: High clearance is a commonly encountered issue in drug discovery. Here we present a centralized metabolic soft spot identification assay with adequate capacity and turnaround time to support the metabolic optimization needs of an entire discovery organization.

Methodology: An integrated quan/qual approach utilizing both an orthogonal sample-pooling methodology and software-assisted structure elucidation was developed to enable the assay. Major metabolic soft spots in liver microsomes (rodent and human) were generated in a batch mode, along with kinetics of parent disappearance and metabolite formation, typically within 1 week of incubation.

Results & conclusion: A centralized metabolic soft spot identification assay has been developed and has successfully impacted discovery project teams in mitigating instability and establishing potential structure-metabolism relationships.

Keywords: dual-concentration incubation; metabolic soft spot; metabolic stability; orthogonal sample pooling; software assisted data processing.

MeSH terms

  • Animals
  • Biological Assay
  • Chromatography, Liquid / standards*
  • Drug Discovery / methods*
  • Humans
  • Kinetics
  • Metabolomics / methods*
  • Mice
  • Microsomes, Liver / metabolism*
  • Rats
  • Software*
  • Tandem Mass Spectrometry / standards*