PMID- 25888743 OWN - NLM STAT- MEDLINE DCOM- 20160119 LR - 20211203 IS - 1479-5876 (Electronic) IS - 1479-5876 (Linking) VI - 13 DP - 2015 Apr 2 TI - Characterization of HIV-1 entry inhibitors with broad activity against R5 and X4 viral strains. PG - 107 LID - 10.1186/s12967-015-0461-9 [doi] LID - 107 AB - BACKGROUND: Combined antiretroviral therapy has drastically reduced mortality and morbidity of HIV-infected individuals. Nevertheless long-term toxicity and appearance of viral resistance hampers the prolonged effectiveness of combination therapy, requiring a continuous input of drugs to replace those utilized in combination regimens. We here investigated the anti-HIV activity of novel derivatives of the suradista chemical class. METHODS: Compounds were tested on acute HIV-1 infection of activated peripheral blood mononuclear cells. HIV production was monitored by enzyme-linked immunosorbent assay measuring the protein p24 released in culture supernatants. Fusion assays were carried out to study the mechanism of action of these compounds. A modified version of a previously established recombinant vaccinia virus-based assay was used measuring activation of a reporter gene upon fusion of two distinct cell populations. Flow cytometry was performed in competition assays for the binding of several antibodies targeting different sites of the viral envelope glycoprotein gp120, or the receptor CD4, or the coreceptors CXCR4 and CCR5. RESULTS: Four compounds inhibited replication of a prototypic R5 (BaL) and X4 (IIIB) laboratory-adapted HIV-1 strain at low micromolar concentrations, in the absence of cytotoxicity. Approximately a ten fold greater activity was achieved against the X4 as compared to the R5 strain. The compounds blocked X4 and R5 HIV-1 fusion, a step of viral entry. This activity appeared specific for HIV-1, as entry of human herpesvirus 6 (HHV-6) and influenza virus was not substantially affected. Further investigation of the inhibitory mechanism revealed that these new molecules target the viral envelope, rather than the coreceptors, as previously shown for a congener of the same class characterized by a long plasmatic half-life. Indeed ND-4043, the most active compound, specifically competed with binding of monoclonal antibodies against the CD4-binding site (CD4-BS) and coreceptor-binding site (CoR-BS) of gp120. These compounds displayed broad anti-HIV activity, as they inhibited various primary R5, X4 and, importantly, dualtropic R5X4 HIV-1 isolates. Of the four derivatives tested, the dimeric compounds were consistently more potent than the monomeric ones. CONCLUSIONS: Given their unique features, these molecules represent promising candidates for further development and exploitation as anti-HIV therapeutics. FAU - Sironi, Francesca AU - Sironi F AD - Unit of Human Virology, Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Hospital, Via Olgettina 60, 20132, Milan, Italy. francesca.sironi@hsr.it. FAU - Malnati, Mauro AU - Malnati M AD - Unit of Human Virology, Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Hospital, Via Olgettina 60, 20132, Milan, Italy. mauro.malnati@hsr.it. FAU - Mongelli, Nicola AU - Mongelli N AD - , Via Tertulliano 38, 20137, Milano, Italy. nivapumo@gmail.com. FAU - Cozzi, Paolo AU - Cozzi P AD - , Via Zanella 48/5, Milano, 20133, Italy. cozzipa@tin.it. FAU - Guzzo, Christina AU - Guzzo C AD - Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), Bethesda, MD, 20892, USA. christina.guzzo@nih.gov. FAU - Ghezzi, Silvia AU - Ghezzi S AD - Unit of Viral Pathogens and Biosafety, Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Hospital, Via Olgettina 60, 20132, Milan, Italy. silvia.ghezzi@hsr.it. FAU - Martinez-Romero, Carles AU - Martinez-Romero C AD - Department of Microbiology, New York, NY, 10029, USA. carles.martinez@mssm.edu. AD - Global Health and Emerging Pathogens Institute, New York, NY, 10029, USA. carles.martinez@mssm.edu. FAU - Garcia-Sastre, Adolfo AU - Garcia-Sastre A AD - Department of Microbiology, New York, NY, 10029, USA. adolfo.garcia-sastre@mssm.edu. AD - Global Health and Emerging Pathogens Institute, New York, NY, 10029, USA. adolfo.garcia-sastre@mssm.edu. AD - Department of Medicine, Division of Infectious Diseases, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA. adolfo.garcia-sastre@mssm.edu. FAU - Lusso, Paolo AU - Lusso P AD - Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), Bethesda, MD, 20892, USA. plusso@niaid.nih.gov. FAU - Jabes, Daniela AU - Jabes D AD - NeED Pharmaceuticals srl, Viale Ortles 22/4, 20139, Milan, Italy. djabes@needpharma.com. FAU - Biswas, Priscilla AU - Biswas P AD - Unit of Molecular Immunology, Division of Genetics and Cell Biology, San Raffaele Hospital, Via Olgettina 60, 20132, Milan, Italy. priscilla.biswas@hsr.it. LA - eng GR - HHSN266200700010C/AI/NIAID NIH HHS/United States GR - HHSN 266200700010C/PHS HHS/United States GR - Intramural NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, N.I.H., Intramural PT - Research Support, Non-U.S. Gov't DEP - 20150402 PL - England TA - J Transl Med JT - Journal of translational medicine JID - 101190741 RN - 0 (Antiviral Agents) RN - 0 (Benzylamines) RN - 0 (Cyclams) RN - 0 (Cyclohexanes) RN - 0 (HIV Envelope Protein gp120) RN - 0 (HIV Fusion Inhibitors) RN - 0 (Heterocyclic Compounds) RN - 0 (Receptors, CCR5) RN - 0 (Receptors, CXCR4) RN - 0 (Triazoles) RN - MD6P741W8A (Maraviroc) RN - S915P5499N (plerixafor) SB - IM MH - 3T3 Cells MH - Animals MH - Antiviral Agents/pharmacology MH - Benzylamines MH - Cell Death/drug effects MH - Cell Line MH - Cyclams MH - Cyclohexanes/pharmacology MH - Flow Cytometry MH - HIV Envelope Protein gp120/metabolism MH - HIV Fusion Inhibitors/*pharmacology MH - HIV Infections/pathology/virology MH - HIV-1/drug effects/*physiology MH - Heterocyclic Compounds/pharmacology MH - Humans MH - Maraviroc MH - Membrane Fusion/drug effects MH - Mice MH - Receptors, CCR5/metabolism MH - Receptors, CXCR4/metabolism MH - Triazoles/pharmacology MH - Virus Internalization/*drug effects MH - Virus Replication/drug effects PMC - PMC4399250 EDAT- 2015/04/19 06:00 MHDA- 2016/01/20 06:00 PMCR- 2015/04/02 CRDT- 2015/04/19 06:00 PHST- 2014/12/15 00:00 [received] PHST- 2015/03/11 00:00 [accepted] PHST- 2015/04/19 06:00 [entrez] PHST- 2015/04/19 06:00 [pubmed] PHST- 2016/01/20 06:00 [medline] PHST- 2015/04/02 00:00 [pmc-release] AID - 10.1186/s12967-015-0461-9 [pii] AID - 461 [pii] AID - 10.1186/s12967-015-0461-9 [doi] PST - epublish SO - J Transl Med. 2015 Apr 2;13:107. doi: 10.1186/s12967-015-0461-9.