Related Experiment Video

Updated: Sep 28, 2025

A Semi-High-Throughput Adaptation of the NADH-Coupled ATPase Assay for Screening Small Molecule Inhibitors
10:28

A Semi-High-Throughput Adaptation of the NADH-Coupled ATPase Assay for Screening Small Molecule Inhibitors

Published on: August 17, 2019

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Discovery of small-molecule inhibitors of RUVBL1/2 ATPase

Gang Zhang1, Feng Wang1, Shan Li1

  • 1Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, United States.

Bioorganic & Medicinal Chemistry
|April 1, 2022
PubMed

Insights

Researchers identified novel pyrazolo[1,5-a]pyrimidine-3-carboxamide compounds targeting RUVBL1/2 ATPases, crucial in cancer progression. Compound 18 demonstrated significant inhibitory activity, offering a promising lead for new anticancer drug development.

Area of Science:

  • Medicinal Chemistry
  • Molecular Biology
  • Oncology

Background:

  • RUVBL1 and RUVBL2 are highly conserved AAA ATPases implicated in cancer progression.
  • These ATPases are attractive targets for developing novel anticancer therapeutics.

Purpose of the Study:

  • To identify compounds with inhibitory activity against the RUVBL1/2 complex using docking-based virtual screening.
  • To synthesize and evaluate pyrazolo[1,5-a]pyrimidine-3-carboxamide analogs as potential anticancer agents.

Main Methods:

  • Docking-based virtual screening to identify initial hits.
  • Synthesis of pyrazolo[1,5-a]pyrimidine-3-carboxamide analogs.
  • Enzymatic and cellular assays to determine inhibitory activity.
  • Structure-activity relationship analysis.
  • Mass spectrometry-based proteomic analysis to explore cellular effects.

Main Results:

  • Seven compounds exhibited inhibitory activity against the RUVBL1/2 complex.
  • Compound 18 showed potent inhibition with IC50 values of 6.0 ± 0.6 μM (RUVBL1/2 complex) and 7.7 ± 0.9 μM (RUVBL1).
  • Compound 18 demonstrated significant IC50 values in various cancer cell lines (A549, H1795, HCT116, MDA-MB-231).
  • Structure-activity relationship studies identified key structural features for inhibitory activity.

Conclusions:

  • Compound 18 is a potent inhibitor of the RUVBL1/2 complex and exhibits anticancer activity in multiple cell lines.
  • The pyrazolo[1,5-a]pyrimidine-3-carboxamide scaffold serves as a promising starting point for developing improved anticancer therapeutics.
  • Further structural modifications of compound 18 are warranted to optimize potency, selectivity, and pharmacokinetic properties.
Keywords:
AAA ATPaseDockingOrganic synthesisPontinProteomicsPyrazolo[1,5-a]pyrimidine-3-carboxamideRUVBL1RUVBL2, Reptin

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