Format

Send to

Choose Destination
ChemMedChem. 2016 Apr 19;11(8):862-9. doi: 10.1002/cmdc.201500526. Epub 2016 Jan 28.

Fragment-Based Protein-Protein Interaction Antagonists of a Viral Dimeric Protease.

Author information

1
Department of Pharmaceutical Chemistry, University of California, San Francisco, CA, 94158-2280, USA.
2
Biophysics Graduate Group, University of California, San Francisco, CA, 94158-2280, USA.
3
Chemistry and Chemical Biology Graduate Group, University of California, San Francisco, CA, 94158-2280, USA.
4
Novartis Institutes for BioMedical Research, Forum 1, Novartis Campus, 4002, Basel, Switzerland.
5
Department of Pharmaceutical Chemistry, University of California, San Francisco, CA, 94158-2280, USA. Charles.Craik@ucsf.edu.

Abstract

Fragment-based drug discovery has shown promise as an approach for challenging targets such as protein-protein interfaces. We developed and applied an activity-based fragment screen against dimeric Kaposi's sarcoma-associated herpesvirus protease (KSHV Pr) using an optimized fluorogenic substrate. Dose-response determination was performed as a confirmation screen, and NMR spectroscopy was used to map fragment inhibitor binding to KSHV Pr. Kinetic assays demonstrated that several initial hits also inhibit human cytomegalovirus protease (HCMV Pr). Binding of these hits to HCMV Pr was also confirmed by NMR spectroscopy. Despite the use of a target-agnostic fragment library, more than 80 % of confirmed hits disrupted dimerization and bound to a previously reported pocket at the dimer interface of KSHV Pr, not to the active site. One class of fragments, an aminothiazole scaffold, was further explored using commercially available analogues. These compounds demonstrated greater than 100-fold improvement of inhibition. This study illustrates the power of fragment-based screening for these challenging enzymatic targets and provides an example of the potential druggability of pockets at protein-protein interfaces.

KEYWORDS:

NMR spectroscopy; dimer disruption; fragment-based screening; human herpesviruses; proteases

PMID:
26822284
PMCID:
PMC4838516
DOI:
10.1002/cmdc.201500526
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Wiley Icon for PubMed Central
Loading ...
Support Center