A covalent chemical probe for Chikungunya nsP2 cysteine protease with antialphaviral activity and proteome-wide selectivity
Chikungunya is a mosquito-borne viral disease that causes fever and severe joint pain for which there is no direct acting drug treatments. Vinyl sulfone SGC-NSP2PRO-1 (3) was identified as a potent inhibitor of the nsP2 cysteine protease (nsP2pro) that reduced viral titer against infectious isolates of Chikungunya and other alphaviruses. The covalent warhead in 3 captured the active site C478 and inactivated nsP2pro with a k<sub>inact</sub>/K<sub>i</sub> ratio of 5950 M<sup>-1</sup> s<sup>-1</sup>. The vinyl sulfone 3 was inactive across a panel of 23 other cysteine proteases and demonstrated remarkable proteome-wide selectivity by two chemoproteomic methods. A negative control analog SGC-NSP2PRO-1N (4) retained the isoxazole core and covalent warhead but demonstrated > 100-fold decrease in enzyme inhibition. Both 3 and 4 were stable across a wide range of pH in solution and upon prolonged storage as solids. Vinyl sulfone 3 and its negative control 4 will find utility as high-quality chemical probes to study the role of the nsP2pro in cellular studies of alphaviral replication and virulence.
Authors
- Kesatebrhan Haile Asressu (ORCID: https://orcid.org/0000-0003-4803-4739)
- Thomas S. Webb
- C.H. Arrowsmith (ORCID: https://orcid.org/0000-0002-4971-3250)
- Daniel N. Streblow (ORCID: https://orcid.org/0000-0002-6828-2492)
- Sumera Perveen (ORCID: https://orcid.org/0000-0002-8898-9540)
- John D. Sears (ORCID: https://orcid.org/0000-0002-4713-2241)
- Zachary J. Streblow
- Mark T. Heise (ORCID: https://orcid.org/0000-0001-5829-8477)
- Kenneth H. Pearce (ORCID: https://orcid.org/0000-0001-8138-5421)
- Anirban Ghoshal (ORCID: https://orcid.org/0000-0001-9359-6074)
- Eric M. Merten (ORCID: https://orcid.org/0000-0002-5306-0790)
- Angie L. Mordant (ORCID: https://orcid.org/0000-0002-0151-4498)
- Sabian A. Martinez (ORCID: https://orcid.org/0000-0002-1333-5083)
- Bennett Davenport (ORCID: https://orcid.org/0000-0001-8155-191X)
- Thomas E. Morrison (ORCID: https://orcid.org/0000-0002-1811-2938)
- Nathaniel J. Moorman (ORCID: https://orcid.org/0000-0003-4952-6858)
- Isabella Law
- Laura E. Herring (ORCID: https://orcid.org/0000-0003-4496-7312)
- Rafael M. Couñago (ORCID: https://orcid.org/0000-0003-1847-5090)
- Aurora Cabrera
- Stefanie Howell (ORCID: https://orcid.org/0000-0002-2039-9245)
- Timothy M. Willson (ORCID: https://orcid.org/0000-0003-4181-8223)
- Mohammad Anwar Hossain (ORCID: https://orcid.org/0000-0001-8684-8755)
- Peter J. Brown (ORCID: https://orcid.org/0000-0002-8454-0367)
- Edwin G. Tse (ORCID: https://orcid.org/0000-0002-9133-9274)
- Jane Burdick (ORCID: https://orcid.org/0009-0001-9719-8302)
- Noah L. Morales
Institutions
- University of North Carolina at Chapel Hill (US)
- Oregon Health & Science University (US)
- Universidade Estadual de Campinas (UNICAMP) (BR)
- University of Toronto (CA)
- Structural Genomics Consortium (CA)
- University of Colorado Denver (US)
Publication Details
- Journal
- Scientific Reports
- Published
- 2025-03-01
- DOI
- https://doi.org/10.1038/s41598-025-91673-x
- Citations
- 8
- Primary Topic
- Mosquito-borne diseases and control
- Type
- article
- Field-Weighted Citation Impact
- 15.03
Funders
- National Institutes of Health