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Chemoproteomic Profiling of Covalent XPO1 Inhibitors to Assess Target Engagement and Selectivity

Jeffrey G Martin1, Jennifer A Ward2,3, Felix Feyertag2,3

  • 1Biogen, Chemical Biology & Proteomics 225 Binney Street, Cambridge, MA 02142, USA.

Chembiochem : a European Journal of Chemical Biology
|April 22, 2021
PubMed

Insights

New clickable probes enable precise measurement of XPO1 inhibitor engagement and selectivity in live cells. These tools aid in understanding efficacy and toxicity for cancer and neurodegenerative disease treatments.

Area of Science:

  • Pharmacology
  • Chemical Biology
  • Molecular Biology

Background:

  • Selinexor, a covalent XPO1 inhibitor, is approved for refractory multiple myeloma.
  • XPO1 inhibitors are under investigation for various hematologic and solid tumor malignancies, glioblastoma multiforme (GBM), and amyotrophic lateral sclerosis (ALS).
  • Assessing target engagement and selectivity of covalent inhibitors is crucial for optimizing efficacy and minimizing toxicity.

Purpose of the Study:

  • To develop clickable probes for labeling XPO1 in live cells using selinexor and eltanexor.
  • To assess target engagement and selectivity of XPO1 inhibitors.
  • To provide tools for interrogating XPO1 biology and inhibition.

Main Methods:

  • Development of clickable probes based on selinexor and eltanexor.
  • Mass spectrometry-based chemoproteomic workflows for proteome-wide selectivity profiling.
  • Thermal profiling analysis for orthogonal measurement of XPO1 engagement.

Main Results:

  • Selinexor and eltanexor probes successfully labeled XPO1 in live cells.
  • Chemoproteomic profiling demonstrated high selectivity of selinexor and eltanexor for XPO1.
  • Thermal profiling confirmed XPO1 engagement by selinexor in live cells.

Conclusions:

  • The developed clickable probes and assays are valuable tools for studying XPO1 inhibition.
  • These tools can advance the understanding of XPO1 biology in cellular and in vivo systems.
  • This research supports the development of targeted therapies for multiple myeloma and other diseases.
Keywords:
XPO1 inhibitorschemical proteomicsclickable probestarget engagement

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