title: A new class of multimerization selective inhibitors of HIV-1 integrase.
abstract: The quinoline-based allosteric HIV-1 integrase (IN) inhibitors (ALLINIs) are promising candidates for clinically useful antiviral agents. Studies using these compounds have highlighted the role of IN in both early and late stages of virus replication. However, dissecting the exact mechanism of action of the quinoline-based ALLINIs has been complicated by the multifunctional nature of these inhibitors because they both inhibit IN binding with its cofactor LEDGF/p75 and promote aberrant IN multimerization with similar potencies in vitro. Here we report design of small molecules that allowed us to probe the role of HIV-1 IN multimerization independently from IN-LEDGF/p75 interactions in infected cells. We altered the rigid quinoline moiety in ALLINIs and designed pyridine-based molecules with a rotatable single bond to allow these compounds to bridge between interacting IN subunits optimally and promote oligomerization. The most potent pyridine-based inhibitor, KF116, potently (EC50 of 0.024 µM) blocked HIV-1 replication by inducing aberrant IN multimerization in virus particles, whereas it was not effective when added to target cells. Furthermore, KF116 inhibited the HIV-1 IN variant with the A128T substitution, which confers resistance to the majority of quinoline-based ALLINIs. A genome-wide HIV-1 integration site analysis demonstrated that addition of KF116 to target or producer cells did not affect LEDGF/p75-dependent HIV-1 integration in host chromosomes, indicating that this compound is not detectably inhibiting IN-LEDGF/p75 binding. These findings delineate the significance of correctly ordered IN structure for HIV-1 particle morphogenesis and demonstrate feasibility of exploiting IN multimerization as a therapeutic target. Furthermore, pyridine-based compounds present a novel class of multimerization selective IN inhibitors as investigational probes for HIV-1 molecular biology.
authors.affiliation: Center for Retrovirus Research and College of Pharmacy, The Ohio State University, Columbus, Ohio, United States of America.
authors.forename: Amit
authors.initials: A
authors.lastname: Sharma
authors.affiliation: Center for Retrovirus Research and College of Pharmacy, The Ohio State University, Columbus, Ohio, United States of America.
authors.forename: Alison
authors.initials: A
authors.lastname: Slaughter
authors.affiliation: Division of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, The Ohio State University, Columbus, Ohio, United States of America.
authors.forename: Nivedita
authors.initials: N
authors.lastname: Jena
authors.affiliation: Center for Retrovirus Research and College of Pharmacy, The Ohio State University, Columbus, Ohio, United States of America.
authors.forename: Lei
authors.initials: L
authors.lastname: Feng
authors.affiliation: Center for Retrovirus Research and College of Pharmacy, The Ohio State University, Columbus, Ohio, United States of America.
authors.forename: Jacques J
authors.initials: JJ
authors.lastname: Kessl
authors.affiliation: Department of Molecular Medicine & Division of Infectious Diseases, Mayo Clinic College of Medicine, Rochester, Minnesota, United States of America.
authors.forename: Hind J
authors.initials: HJ
authors.lastname: Fadel
authors.affiliation: Perelman School of Medicine, Department of Microbiology, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.
authors.forename: Nirav
authors.initials: N
authors.lastname: Malani
authors.affiliation: Perelman School of Medicine, Department of Microbiology, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.
authors.forename: Frances
authors.initials: F
authors.lastname: Male
authors.affiliation: Center for Retrovirus Research and Department of Veterinary Biosciences, The Ohio State University, Columbus, Ohio, United States of America.
authors.forename: Li
authors.initials: L
authors.lastname: Wu
authors.affiliation: Department of Molecular Medicine & Division of Infectious Diseases, Mayo Clinic College of Medicine, Rochester, Minnesota, United States of America.
authors.forename: Eric
authors.initials: E
authors.lastname: Poeschla
authors.affiliation: Perelman School of Medicine, Department of Microbiology, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.
authors.forename: Frederic D
authors.initials: FD
authors.lastname: Bushman
authors.affiliation: Division of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, The Ohio State University, Columbus, Ohio, United States of America.
authors.forename: James R
authors.initials: JR
authors.lastname: Fuchs
authors.affiliation: Center for Retrovirus Research and College of Pharmacy, The Ohio State University, Columbus, Ohio, United States of America.
authors.forename: Mamuka
authors.initials: M
authors.lastname: Kvaratskhelia
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D019427:HIV Integrase / Q000378:metabolism*; D019428:HIV Integrase Inhibitors / Q000494:pharmacology*; D015497:HIV-1 / Q000187:drug effects; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology*; D011485:Protein Binding / Q000187:drug effects; D055503:Protein Multimerization / Q000187:drug effects*; D011804:Quinolines / Q000494:pharmacology; D016662:Virus Integration / Q000187:drug effects*; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D019428:HIV Integrase Inhibitors; D036341:Intercellular Signaling Peptides and Proteins; D011804:Quinolines; C119380:lens epithelium-derived growth factor; C037219:quinoline; D019427:HIV Integrase; C514801:p31 integrase protein, Human immunodeficiency virus 1