(Thia)calixarenephosphonic Acids as Potent Inhibitors of the Nucleic Acid Chaperone Activity of the HIV-1 Nucleocapsid Protein with a New Binding Mode and Multitarget Antiviral Activity

Nicolas Humbert (1) , Lesia Kovalenko (1) , Francesco Saladini , Alessia Giannini , Manuel Pires (1) , Thomas Botzanowski (2) , Sergiy Cherenok , Christian Boudier (1) , Kamal Sharma (1) , Eleonore Real (1) , Olga Zaporozhets (3) , Sarah Cianférani (2) , Carole Seguin-Devaux (4) , Federica Poggialini (5) , Maurizio Botta (5) , Maurizio Zazzi (5) , Vitaly Kalchenko (6) , Mattia Mori (5) , Yves Mély (1)

1 LBP - Laboratoire de Bioimagerie et Pathologies
2 DSA-IPHC - Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires
3 National Taras Shevchenko University of Kiev
4 Centre de Recherche Public de la Santé
5 UNISI - Università degli Studi di Siena = University of Siena
6 Institute of Organic Chemistry of NASU [Kyiv]

The nucleocapsid protein (NC) is a highly conserved protein that plays key roles in HIV-1 replication through its nucleic acid chaperone properties mediated by its two zinc fingers and basic residues. NC is a promising target for antiviral therapy, particularly to control viral strains resistant to currently available drugs. Since calixarenes with antiviral properties have been described, we explored the ability of calixarene hydroxymethylphosphonic or sulfonic acids to inhibit NC chaperone properties and exhibit antiviral activity. By using fluorescence-based assays, we selected four calixarenes inhibiting NC chaperone activity with submicromolar IC50 values. These compounds were further shown by mass spectrometry, isothermal titration calorimetry, and fluorescence anisotropy to bind NC with no zinc ejection and to compete with nucleic acids for the binding to NC. Molecular dynamic simulations further indicated that these compounds interact via their phosphonate or sulfonate groups with the basic surface of NC but not with the hydrophobic plateau at the top of the folded fingers. Cellular studies showed that the most soluble compound CIP201 inhibited the infectivity of wild-type and drug-resistant HIV-1 strains at low micromolar concentrations, primarily targeting the early steps of HIV-1 replication. Moreover, CIP201 was also found to inhibit the flipping and polymerization activity of reverse transcriptase. Calixarenes thus form a class of noncovalent NC inhibitors, endowed with a new binding mode and multitarget antiviral activity.

Mots clés: calixarenes; NC inhibitors; fluorescence; HIV-1; nucleocapsid protein

Domaines: Chimie

Nicolas Humbert, Lesia Kovalenko, Francesco Saladini, Alessia Giannini, Manuel Pires, et al.. (Thia)calixarenephosphonic Acids as Potent Inhibitors of the Nucleic Acid Chaperone Activity of the HIV-1 Nucleocapsid Protein with a New Binding Mode and Multitarget Antiviral Activity. ACS Infectious Diseases, 2020, ⟨10.1021/acsinfecdis.9b00290⟩. ⟨hal-02493003⟩