Pan-serotype dengue virus inhibitor JNJ-A07 targets NS4A-2K-NS4B interaction with NS2B/NS3 and blocks replication organelle formation

  • Nat Commun. 2024 Jul 19;15(1):6080. doi: 10.1038/s41467-024-50437-3.
Dominik Kiemel  1 ,  Ann-Sophie Helene Kroell  1 ,  Solène Denolly  1 ,  Uta Haselmann  1 ,  Jean-François Bonfanti  2  3 ,  Jose Ignacio Andres  4 ,  Brahma Ghosh  5 ,  Peggy Geluykens  6 ,  Suzanne J F Kaptein  7 ,  Lucas Wilken  8 ,  Pietro Scaturro  8 ,  Johan Neyts  7 ,  Marnix Van Loock  9 ,  Olivia Goethals  9 ,  Ralf Bartenschlager  10  11
Affiliations
  • 1. Heidelberg University, Medical Faculty Heidelberg, Department of Infectious Diseases, Molecular Virology, Center for Integrative Infectious Disease Research, Heidelberg, Germany.
  • 2. Janssen Infectious Diseases Discovery, Janssen-Cilag, Val de Reuil, France.
  • 3. Evotec, Toulouse, France.
  • 4. Discovery Chemistry, Janssen R&D, a Johnson & Johnson company, Toledo, Spain.
  • 5. Discovery Chemistry, Janssen R&D, a Johnson & Johnson company, Spring House, PA, USA.
  • 6. Discovery, Charles River Beerse, Beerse, Belgium.
  • 7. Department of Microbiology, Immunology and Transplantation, Rega Institute for Medical Research, Laboratory of Virology and Chemotherapy, KU Leuven, Leuven, Belgium.
  • 8. Leibniz Institute of Virology, Hamburg, Germany.
  • 9. Janssen Global Public Health, Janssen Pharmaceutica NV, a Johnson & Johnson company, Beerse, Belgium.
  • 10. Heidelberg University, Medical Faculty Heidelberg, Department of Infectious Diseases, Molecular Virology, Center for Integrative Infectious Disease Research, Heidelberg, Germany. [email protected].
  • 11. German Centre for Infection Research, Heidelberg partner site, Heidelberg, Germany. [email protected].
PMID: 39030239 DOI: 10.1038/s41467-024-50437-3
Abstract

Dengue fever represents a significant medical and socio-economic burden in (sub)tropical regions, yet antivirals for treatment or prophylaxis are lacking. JNJ-A07 was described as highly active against the different genotypes within each serotype of the disease-causing Dengue Virus (DENV). Based on clustering of resistance mutations it has been assumed to target DENV non-structural protein 4B (NS4B). Using a photoaffinity labeling compound with high structural similarity to JNJ-A07, here we demonstrate binding to NS4B and its precursor NS4A-2K-NS4B. Consistently, we report recruitment of the compound to intracellular sites enriched for these proteins. We further specify the mechanism-of-action of JNJ-A07, which has virtually no effect on viral polyprotein cleavage, but targets the interaction between the NS2B/NS3 protease/helicase complex and the NS4A-2K-NS4B cleavage intermediate. This interaction is functionally linked to de novo formation of vesicle packets (VPs), the sites of DENV RNA replication. JNJ-A07 blocks VPs biogenesis with little effect on established ones. A similar mechanism-of-action was found for another NS4B inhibitor, NITD-688. In summary, we unravel the Antiviral mechanism of these NS4B-targeting molecules and show how DENV employs a short-lived cleavage intermediate to carry out an early step of the viral life cycle.