title: Isolation and characterization of novel human immunodeficiency virus integrase inhibitors from fungal metabolites.
abstract: We have identified a series of novel inhibitors of human immunodeficiency virus type 1 (HIV-1) integrase by randomly screening natural product extracts using an in vitro biochemical assay designed to identify inhibitors of integrase-catalysed strand transfer. Equisetin recovered from the fungus Fusarium heterosporum and a novel enantiomeric homologue of equisetin from Phoma sp. were isolated as inhibitors of HIV-1 integrase in vitro. Two additional analogues, a novel decalin derivative, integric acid, and oteromycin were also discovered to be inhibitors of integrase. Equisetin and related compounds inhibit 3' end-processing and strand transfer as well as disintegration catalysed by either the full-length enzyme or the truncated integrase core domain (amino acids 50-212). These compounds also inhibit strand transfer reactions catalysed by stable complexes assembled in vitro and integration reactions catalysed by pre-integration complexes isolated from HIV-1-infected cells. The compounds described in this report are structurally novel and mechanistically distinct from many previously described inhibitors of HIV-1 integrase. These results demonstrate the utility of using an appropriately configured assay to identify compounds that are effective post-assembly and the potential of isolating novel integrase inhibitors from complex natural product extracts.
authors.affiliation: Department of Antiviral Research, Merck Research Laboratories, West Point, PA 19486, USA. daria_hazuda@merck.com
authors.forename: D
authors.initials: D
authors.lastname: Hazuda
authors.forename: C U
authors.initials: CU
authors.lastname: Blau
authors.forename: P
authors.initials: P
authors.lastname: Felock
authors.forename: J
authors.initials: J
authors.lastname: Hastings
authors.forename: B
authors.initials: B
authors.lastname: Pramanik
authors.forename: A
authors.initials: A
authors.lastname: Wolfe
authors.forename: F
authors.initials: F
authors.lastname: Bushman
authors.forename: C
authors.initials: C
authors.lastname: Farnet
authors.forename: M
authors.initials: M
authors.lastname: Goetz
authors.forename: M
authors.initials: M
authors.lastname: Williams
authors.forename: K
authors.initials: K
authors.lastname: Silverman
authors.forename: R
authors.initials: R
authors.lastname: Lingham
authors.forename: S
authors.initials: S
authors.lastname: Singh
mesh_terms: D001483:Base Sequence; D002264:Carboxylic Acids / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D002384:Catalysis; D017931:DNA Primers; D005670:Fusarium / Q000737:chemistry / Q000378:metabolism*; D019427:HIV Integrase / Q000737:chemistry / Q000302:isolation & purification; D019428:HIV Integrase Inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D009281:Naphthalenes / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D011758:Pyrroles / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D011759:Pyrrolidines / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D011760:Pyrrolidinones / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification; D013764:Tetrahydronaphthalenes
chemical_list: D002264:Carboxylic Acids; D017931:DNA Primers; D019428:HIV Integrase Inhibitors; D009281:Naphthalenes; D011758:Pyrroles; D011759:Pyrrolidines; D011760:Pyrrolidinones; D011994:Recombinant Proteins; D013764:Tetrahydronaphthalenes; C120097:integric acid; C120096:phomasetin; C120095:oteromycin; C024509:equisetin; D019427:HIV Integrase