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Related Experiment Video

Updated: Sep 9, 2025

Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays
13:22

Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays

Published on: October 23, 2019

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IP6 Kinase Inhibitor Screening Assays.

Huijun Wei1,2, James C Barrow3,4

  • 1Department of Pharmacology and Molecular Sciences, Johns Hopkins School of Medicine, Baltimore, MD, USA.

Methods in Molecular Biology (Clifton, N.J.)
|August 29, 2025
PubMed
Summary

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Researchers identified novel inhibitors for IP6K, an enzyme linked to diabetes and thrombosis. Assays confirmed these compounds reduce IP7 levels and bind to IP6K in cells, paving the way for new therapies.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Pharmacology

Background:

  • Inositol pyrophosphates, including 5-diphosphoinositol pentakisphosphate (IP7), are vital in cellular signaling.
  • Inhibition of inositol hexakisphosphate kinase (IP6K) shows therapeutic promise for type II diabetes, thrombosis, kidney disease, and psychiatric disorders.

Purpose of the Study:

  • To describe methods for identifying and validating IP6K inhibitors.
  • To establish robust assays for evaluating IP6K inhibitor efficacy and cellular engagement.

Main Methods:

  • Kinase-activity-based high-throughput screening and dose-response assays for inhibitor identification.
  • Cell-based assays to measure IP7 production in HCT116 cells.
  • Cellular thermal shift assays (CETSAs) to confirm in-cell binding of inhibitors to IP6K.
Keywords:
ADP-GloCellular thermal shiftHiBitHigh-throughput assayIP7Inositol hexakisphosphate kinase

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Main Results:

  • Successful identification of IP6K inhibitors using biochemical assays.
  • Demonstration of IP6K inhibitor-mediated reduction of IP7 levels in HCT116 cells.
  • Confirmation of direct binding of IP6K inhibitors to the target enzyme within cells via CETSAs.

Conclusions:

  • Established a suite of assays for the discovery and characterization of IP6K inhibitors.
  • These methods provide a strong foundation for advancing IP6K inhibitors for therapeutic development in various diseases.