Rapid Optimization Enabled by Single-Molecule Tracking: Discovery of a Potent RUVBL1/2 Inhibitor to Evaluate the Targeting of MYC-Driven Cancers
RuvB-like 1 (RUVBL1) and RuvB-like 2 (RUVBL2) are AAA ATPases that form hetero-oligomeric complexes involved in diverse cellular functions. Increasing evidence implicates the RUVBL1/2 complex as an essential cofactor of MYC, with RUVBL1/2 inhibition reducing c-MYC levels in vitro. Herein, we report a potent RUVBL1/2 inhibitor discovered through a Single-Molecule Tracking (SMT)-driven SAR campaign. Compared with a biochemical ADP-Glo assay, which exhibited limited dynamic range and poor reproducibility under our experimental conditions, the live-cell high-throughput RUVBL SMT assay provided robust and reproducible potency measurements and correlated strongly with cell viability and MYC degradation. Multiparameter optimization yielded compound 18, which demonstrated improved efficacy in a MYC-dependent Burkitt lymphoma xenograft model at a significantly lower dose than the RUVBL1/2 inhibitor CB-6644. This work establishes SMT as a powerful tool to facilitate the drug discovery SAR campaigns and evaluates the therapeutic potential of RUVBL1/2 inhibition in MYC-dependent cancers.
Authors
- Taylor Heuer
- Marcus P. Kelly (ORCID: https://orcid.org/0000-0002-1276-3107)
- Leah Cleary
- Abhijit Tarafder (ORCID: https://orcid.org/0000-0001-5885-0348)
- Jesse D. Vargas (ORCID: https://orcid.org/0000-0002-3671-6778)
- Yangzhong Tang (ORCID: https://orcid.org/0000-0001-7086-0063)
- Grzegorz Nawrocki (ORCID: https://orcid.org/0000-0002-7115-5857)
- Charmaine Fong
- Arnold Tao
- David W. Piotrowski (ORCID: https://orcid.org/0000-0002-4659-6300)
- Patric Schyman (ORCID: https://orcid.org/0000-0002-5739-3891)
- Huntly M. Morrison (ORCID: https://orcid.org/0000-0003-0091-2395)
- Hilary P. Beck
- Amine Driouchi (ORCID: https://orcid.org/0000-0002-2940-1161)
- Daniel J. Anderson (ORCID: https://orcid.org/0000-0002-3249-2439)
- Jennifer M. Le (ORCID: https://orcid.org/0000-0003-0945-9830)
- Melissa Dumble
- Stephen Jones (ORCID: https://orcid.org/0000-0003-0785-7157)
- William S. R. Forrest (ORCID: https://orcid.org/0000-0003-0985-1453)
- Xiao Chang (ORCID: https://orcid.org/0009-0001-2727-4410)
- Eugene Park (ORCID: https://orcid.org/0000-0001-8432-5168)
- Helen Wong (ORCID: https://orcid.org/0000-0002-3927-8953)
- J.M. Ortega (ORCID: https://orcid.org/0000-0003-4816-4883)
- Ali Tabatabaei
- Lakshmi Akella
- Rohit Malik
- Zachary Howard
- Daniel Bracho
- Quan Zheng
- Kayla VanBuren
- Mai K. Nguyen
- Fernando Rodríguez Pérez
- Qian Du
- Michael Farley (ORCID: https://orcid.org/0009-0004-8775-758X)
- Jason Lenihan
- Liam A. Elliott
- Maité B. Silva
- Xin Zhou
- Samuel T. Albright
- Daisy Ding
- Li Zheng (ORCID: https://orcid.org/0000-0002-4228-7797)
Institutions
- Rapt Therapeutics (United States) (US)
Publication Details
- Journal
- Journal of Medicinal Chemistry
- Published
- 2026-04-22
- DOI
- https://doi.org/10.1021/acs.jmedchem.5c03692
- Primary Topic
- Protein Degradation and Inhibitors
- Type
- article
- Field-Weighted Citation Impact
- 0.00