TY - JOUR
T1 - (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
AU - Humbert, Nicolas
AU - Kovalenko, Lesia
AU - Saladini, Francesco
AU - Giannini, Alessia
AU - Pires, Manuel
AU - Botzanowski, Thomas
AU - Cherenok, Sergiy
AU - Boudier, Christian
AU - Sharma, Kamal K.
AU - Real, Eleonore
AU - Zaporozhets, Olga A.
AU - Cianférani, Sarah
AU - Seguin-Devaux, Carole
AU - Poggialini, Federica
AU - Botta, Maurizio
AU - Zazzi, Maurizio
AU - Kalchenko, Vitaly I.
AU - Mori, Mattia
AU - Mély, Yves
N1 - Funding Information:
This work was supported by European Project THINPAD “Targeting the HIV-1 Nucleocapsid Protein to Fight Antiretroviral Drug Resistance”, FP7 grant agreement 601969, the Agence Nationale de la Recherche (ANR), the French Proteomic Infrastructure (ProFI, ANR-10-INBS-08-03), the French Infrastructure for Integrated Structural Biology (FRISBI) ANR-10-INSB-05 and Instruct-ERIC, and a Ph.D. fellowship from Institute de Recherche Servier to T.B. Y.M. is grateful to the Institut Universitaire de France (IUF) for support and providing additional time for research.
Publisher Copyright:
© 2020 American Chemical Society.
PY - 2020/4/10
Y1 - 2020/4/10
N2 - 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.
AB - 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.
KW - HIV-1
KW - NC inhibitors
KW - calixarenes
KW - fluorescence
KW - nucleocapsid protein
UR - http://www.scopus.com/inward/record.url?scp=85080946616&partnerID=8YFLogxK
U2 - 10.1021/acsinfecdis.9b00290
DO - 10.1021/acsinfecdis.9b00290
M3 - Article
C2 - 32045204
AN - SCOPUS:85080946616
SN - 2373-8227
VL - 6
SP - 687
EP - 702
JO - ACS Infectious Diseases
JF - ACS Infectious Diseases
IS - 4
ER -