title: Potent inhibition of HIV-1 replication by novel non-peptidyl small molecule inhibitors of protease dimerization.
abstract: Dimerization of HIV-1 protease subunits is essential for its proteolytic activity, which plays a critical role in HIV-1 replication. Hence, the inhibition of protease dimerization represents a unique target for potential intervention of HIV-1. We developed an intermolecular fluorescence resonance energy transfer-based HIV-1-expression assay employing cyan and yellow fluorescent protein-tagged protease monomers. Using this assay, we identified non-peptidyl small molecule inhibitors of protease dimerization. These inhibitors, including darunavir and two experimental protease inhibitors, blocked protease dimerization at concentrations of as low as 0.01 microm and blocked HIV-1 replication with IC(50) values of 0.0002-0.48 microm. These agents also inhibited the proteolytic activity of mature protease. Other approved anti-HIV-1 agents examined except tipranavir, a CCR5 inhibitor, and soluble CD4 failed to block the dimerization event. Once protease monomers dimerize to become mature protease, mature protease is not dissociated by this dimerization inhibition mechanism, suggesting that these agents block dimerization at the nascent stage of protease maturation. The proteolytic activity of mature protease that managed to undergo dimerization despite the presence of these agents is likely to be inhibited by the same agents acting as conventional protease inhibitors. Such a dual inhibition mechanism should lead to highly potent inhibition of HIV-1.
authors.affiliation: Department of Hematology, Kumamoto University Graduate School of Medical and Pharmaceutical Sciences, 1-1-1 Honjo, Kumamoto 860-8556, Japan; Department of Infectious Diseases, Kumamoto University Graduate School of Medical and Pharmaceutical Sciences, 1-1-1 Honjo, Kumamoto 860-8556, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Koh
authors.affiliation: Department of Hematology, Kumamoto University Graduate School of Medical and Pharmaceutical Sciences, 1-1-1 Honjo, Kumamoto 860-8556, Japan; Department of Infectious Diseases, Kumamoto University Graduate School of Medical and Pharmaceutical Sciences, 1-1-1 Honjo, Kumamoto 860-8556, Japan.
authors.forename: Shintaro
authors.initials: S
authors.lastname: Matsumi
authors.affiliation: Experimental Retrovirology Section, NCI, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: Debananda
authors.initials: D
authors.lastname: Das
authors.affiliation: Department of Hematology, Kumamoto University Graduate School of Medical and Pharmaceutical Sciences, 1-1-1 Honjo, Kumamoto 860-8556, Japan; Department of Infectious Diseases, Kumamoto University Graduate School of Medical and Pharmaceutical Sciences, 1-1-1 Honjo, Kumamoto 860-8556, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Amano
authors.affiliation: Retroviral Disease Section, HIV and AIDS Malignancy Branch, NCI, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: David A
authors.initials: DA
authors.lastname: Davis
authors.affiliation: Departments of Chemistry and Medicinal Chemistry, Purdue University, West Lafayette, Indiana 47907.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Li
authors.affiliation: Departments of Chemistry and Medicinal Chemistry, Purdue University, West Lafayette, Indiana 47907.
authors.forename: Sofiya
authors.initials: S
authors.lastname: Leschenko
authors.affiliation: Departments of Chemistry and Medicinal Chemistry, Purdue University, West Lafayette, Indiana 47907.
authors.forename: Abigail
authors.initials: A
authors.lastname: Baldridge
authors.affiliation: Department of Viral Infections, Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan.
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Shioda
authors.affiliation: Retroviral Disease Section, HIV and AIDS Malignancy Branch, NCI, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: Robert
authors.initials: R
authors.lastname: Yarchoan
authors.affiliation: Departments of Chemistry and Medicinal Chemistry, Purdue University, West Lafayette, Indiana 47907.
authors.forename: Arun K
authors.initials: AK
authors.lastname: Ghosh
authors.affiliation: Department of Hematology, Kumamoto University Graduate School of Medical and Pharmaceutical Sciences, 1-1-1 Honjo, Kumamoto 860-8556, Japan; Department of Infectious Diseases, Kumamoto University Graduate School of Medical and Pharmaceutical Sciences, 1-1-1 Honjo, Kumamoto 860-8556, Japan; Experimental Retrovirology Section, NCI, National Institutes of Health, Bethesda, Maryland 20892. Electronic address: hmitsuya@helix.nih.gov.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Mitsuya
mesh_terms: D000818:Animals; D065100:CCR5 Receptor Antagonists; D015704:CD4 Antigens / Q000494:pharmacology; D019556:COS Cells; D002522:Chlorocebus aethiops; D000069454:Darunavir; D019281:Dimerization; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D031541:Fluorescence Resonance Energy Transfer; D016333:HIV Protease / Q000737:chemistry / Q000378:metabolism*; D017320:HIV Protease Inhibitors / Q000494:pharmacology*; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D011499:Protein Processing, Post-Translational / Q000187:drug effects* / Q000502:physiology; D011725:Pyridines / Q000494:pharmacology*; D011753:Pyrones / Q000494:pharmacology*; D019713:Receptors, CCR5 / Q000378:metabolism; D013449:Sulfonamides / Q000494:pharmacology*; D014779:Virus Replication / Q000187:drug effects* / Q000502:physiology
chemical_list: D065100:CCR5 Receptor Antagonists; D015704:CD4 Antigens; D017320:HIV Protease Inhibitors; D011725:Pyridines; D011753:Pyrones; D019713:Receptors, CCR5; D013449:Sulfonamides; D016333:HIV Protease; D000069454:Darunavir; C107201:tipranavir