Fragment-based and structure-guided discovery of perforin inhibitors

Jiney Jose (Leading Author), Ruby H.P. Law (Leading Author), Eleanor W.W. Leung, Dorothy C.C. Wai, Hedieh Akhlaghi, Indu R. Chandrashekaran, Tom T. Caradoc-Davies, Ilia Voskoboinik, John Feutrill, David Middlemiss, Devadharshini Jeevarajah, Tanya Bashtannyk-Puhalovich, Anna C. Giddens, Tet Woo Lee, Stephen M.F. Jamieson, Joseph A. Trapani, James C. Whisstock (Leading Author), Julie A. Spicer (Leading Author), Raymond S. Norton (Leading Author)

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Perforin is a pore-forming protein whose normal function enables cytotoxic T and natural killer (NK) cells to kill virus-infected and transformed cells. Conversely, unwanted perforin activity can also result in auto-immune attack, graft rejection and aberrant responses to pathogens. Perforin is critical for the function of the granule exocytosis cell death pathway and is therefore a target for drug development. In this study, by screening a fragment library using NMR and surface plasmon resonance, we identified 4,4-diaminodiphenyl sulfone (dapsone) as a perforin ligand. We also found that dapsone has modest (mM) inhibitory activity of perforin lytic activity in a red blood cell lysis assay in vitro. Sequential modification of this lead fragment, guided by structural knowledge of the ligand binding site and binding pose, and supported by SPR and ligand-detected 19F NMR, enabled the design of nanomolar inhibitors of the cytolytic activity of intact NK cells against various tumour cell targets. Interestingly, the ligands we developed were largely inert with respect to direct perforin-mediated red blood cell lysis but were very potent in the context of perforin's action on delivering granzymes in the immune synapse, the context in which it functions physiologically. Our work indicates that a fragment-based, structure-guided drug discovery strategy can be used to identify novel ligands that bind perforin. Moreover, these molecules have superior physicochemical properties and solubility compared to previous generations of perforin ligands.

Original languageEnglish
Article number115786
Number of pages19
JournalEuropean Journal of Medicinal Chemistry
Volume261
DOIs
Publication statusPublished - 5 Dec 2023

Keywords

  • 4,4-Diaminodiphenyl sulfone
  • Fragment-based drug discovery
  • Natural killer cells
  • Perforin
  • X-ray crystallography

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