SLAS Discovery (Jan 2022)

Using measures of metabolic flux to align screening and clinical development: Avoiding pitfalls to enable translational studies

  • Santhosh Satapati,
  • Daniel P. Downes,
  • Daniel Metzger,
  • Harish Shankaran,
  • Saswata Talukdar,
  • Yingjiang Zhou,
  • Zhao Ren,
  • Michelle Chen,
  • Yeon-Hee Lim,
  • Nathan G. Hatcher,
  • Xiujuan Wen,
  • Payal R. Sheth,
  • David G. McLaren,
  • Stephen F. Previs

Journal volume & issue
Vol. 27, no. 1
pp. 20 – 28

Abstract

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Screening campaigns, especially those aimed at modulating enzyme activity, often rely on measuring substrate→product conversions. Unfortunately, the presence of endogenous substrates and/or products can limit one's ability to measure conversions. As well, coupled detection systems, often used to facilitate optical readouts, are subject to interference. Stable isotope labeled substrates can overcome background contamination and yield a direct readout of enzyme activity. Not only can isotope kinetic assays enable early screening, but they can also be used to follow hit progression in translational (pre)clinical studies. Herein, we consider a case study surrounding lipid biology to exemplify how metabolic flux analyses can connect stages of drug development, caveats are highlighted to ensure reliable data interpretations. For example, when measuring enzyme activity in early biochemical screening it may be enough to quantify the formation of a labeled product. In contrast, cell-based and in vivo studies must account for variable exposure to a labeled substrate (or precursor) which occurs via tracer dilution and/or isotopic exchange. Strategies are discussed to correct for these complications. We believe that measures of metabolic flux can help connect structure-activity relationships with pharmacodynamic mechanisms of action and determine whether mechanistically differentiated biophysical interactions lead to physiologically relevant outcomes. Adoption of this logic may allow research programs to (i) build a critical bridge between primary screening and (pre)clinical development, (ii) elucidate biology in parallel with screening and (iii) suggest a strategy aimed at in vivo biomarker development.

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