Inhibition of respiratory syncytial virus fusion by the small molecule VP-14637 via specific interactions with F protein

Janet Douglas, Marites L. Panis, Edmund Ho, Kuei Ying Lin, Steve H. Krawczyk, Deborah M. Grant, Ruby Cai, Swami Swaminathan, Tomas Cihlar

Research output: Contribution to journalArticle

91 Citations (Scopus)

Abstract

Human respiratory syncytial virus (RSV) is a major cause of respiratory tract infections worldwide. Several novel small-molecule inhibitors of RSV have been identified, but they are still in preclinical or early clinical evaluation. One such inhibitor is a recently discovered triphenol-based molecule, VP-14637 (ViroPharma). Initial experiments suggested that VP-14637 acted early and might be an RSV fusion inhibitor. Here we present studies demonstrating that VP-14637 does not block RSV adsorption but inhibits RSV-induced cell-cell fusion and binds specifically to RSV-infected cells with an affinity corresponding to its inhibitory potency. VP-14637 is capable of specifically interacting with the RSV fusion protein expressed by a T7 vaccinia virus system. RSV variants resistant to VP-14637 were selected; they had mutations localized to two distinct regions of the RSV F protein, heptad repeat 2 (HR2) and the intervening domain between heptad repeat 1 (HR1) and HR2. No mutations arose in HR1, suggesting a mechanism other than direct disruption of the heptad repeat interaction. The F proteins containing the resistance mutations exhibited greatly reduced binding of VP-14637. Despite segregating with the membrane fraction following incubation with intact RSV-infected cells, the compound did not bind to membranes isolated from RSV-infected cells. In addition, binding of VP-14637 was substantially compromised at temperatures of ≤22°C. Therefore, we propose that VP-14637 inhibits RSV through a novel mechanism involving an interaction between the compound and a transient conformation of the RSV F protein.

Original languageEnglish (US)
Pages (from-to)5054-5064
Number of pages11
JournalJournal of Virology
Volume77
Issue number9
DOIs
StatePublished - May 2003
Externally publishedYes

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Respiratory Syncytial Viruses
viruses
Proteins
proteins
mutation
Mutation
VP 14637
viral fusion proteins
Human respiratory syncytial virus
cells
Viral Fusion Proteins
cell fusion
Vaccinia virus
Membranes
Cell Fusion
respiratory tract diseases
Respiratory Tract Infections
Adsorption
adsorption

ASJC Scopus subject areas

  • Immunology

Cite this

Inhibition of respiratory syncytial virus fusion by the small molecule VP-14637 via specific interactions with F protein. / Douglas, Janet; Panis, Marites L.; Ho, Edmund; Lin, Kuei Ying; Krawczyk, Steve H.; Grant, Deborah M.; Cai, Ruby; Swaminathan, Swami; Cihlar, Tomas.

In: Journal of Virology, Vol. 77, No. 9, 05.2003, p. 5054-5064.

Research output: Contribution to journalArticle

Douglas, J, Panis, ML, Ho, E, Lin, KY, Krawczyk, SH, Grant, DM, Cai, R, Swaminathan, S & Cihlar, T 2003, 'Inhibition of respiratory syncytial virus fusion by the small molecule VP-14637 via specific interactions with F protein', Journal of Virology, vol. 77, no. 9, pp. 5054-5064. https://doi.org/10.1128/JVI.77.9.5054-5064.2003
Douglas, Janet ; Panis, Marites L. ; Ho, Edmund ; Lin, Kuei Ying ; Krawczyk, Steve H. ; Grant, Deborah M. ; Cai, Ruby ; Swaminathan, Swami ; Cihlar, Tomas. / Inhibition of respiratory syncytial virus fusion by the small molecule VP-14637 via specific interactions with F protein. In: Journal of Virology. 2003 ; Vol. 77, No. 9. pp. 5054-5064.
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