Inhibition of a viral enzyme by a small-molecule dimer disruptor

Abstract

Small molecule dimer disruptors that inhibit an essential dimeric protease of human Kaposi’s sarcoma-associated herpesvirus (KSHV) were identified by screening an α-helical mimetic library. Subsequently, a second generation of low micromolar inhibitors with improved potency and solubility was synthesized. Complementary methods including size exclusion chromatography and 1H-13C HSQC titration using selectively labeled 13C-Met samples revealed that monomeric protease is enriched in the presence of inhibitor. 1H-15N-HSQC titration studies mapped the inhibitor binding-site to the dimer interface, and mutagenesis studies targeting this region were consistent with a mechanism where inhibitor binding prevents dimerization through the conformational selection of a dynamic intermediate. These results validate the interface of herpesvirus proteases and other similar oligomeric interactions as suitable targets for the development of small molecule inhibitors.

Deciphering the Xcp Pseudomonas aeruginosa Type II Secretion Machinery through Multiple Interactions with Substrates

“…Collectively, our results provide an improved understanding of two important steps of the secretion process: substrate recognition and transport by the machinery. We propose an improved model of the type II secretion process, opening new routes for academic investigation and for antimicrobial targeting utilizing organic disruptors (31).…”

Potential pharmacological chaperones targeting cancer-associated MCL-1 and Parkinson disease-associated α-synuclein

“…Indeed, there are only a few reports on the use of HTS to identify α-helix mimetic small molecules (39)(40)(41). The most frequently used method to identify α-helix mimetics is a competitive binding assay, using a fluorescence polarization (FP).…”

Identification of a Varicella-Zoster Virus Replication Inhibitor That Blocks Capsid Assembly by Interacting with the Floor Domain of the Major Capsid Protein

“…Some derivatives of [2,3-d]oxazinone inhibit the protease activity of the scaffold proteins of HSV, VZV, and CMV (11,59). Dimerization of scaffold proteins is required for the protease activity; compounds that blocked their dimerization were recently reported for HHV-8 (42). Because 35B2 is the first antiviral that targets the conserved herpesvirus MCP, its novel antiviral mechanism can be exploited to target other herpesviruses.…”