title: Substrate-induced stable enzyme-inhibitor complex formation allows tight binding of novel 2-aminopyrimidin-4(3H)-ones to drug-resistant HIV-1 reverse transcriptase mutants.
abstract: We recently reported the synthesis and biological evaluation of a novel series of 5-alkyl-2-(N,N-disubstituted)amino-6-(2,6-difluorophenylalkyl)-3,4-dihydropyrimidin-4(3H)-ones (F(2)-N,N-DABOs). These compounds are highly active against both wild-type HIV-1 and the K103N, Y181C, and Y188L mutant strains. Herein we present novel 6-(2-chloro-6-fluorophenylalkyl)-N,N-DABO (2-Cl-6-F-N,N-DABO) derivatives and investigate the molecular basis for their high-affinity binding to HIV-1 reverse transcriptase (RT). Our results show that the new compounds display higher association rates than the difluoro derivatives toward wild-type HIV-1 RT or drug-resistant RT mutant forms. We also show that they preferentially associate to either the free enzyme or the enzyme-nucleic acid binary complex, and that this binding is stabilized upon formation of the ternary complex between HIV-1 RT and both the nucleic acid and nucleotide substrates. Interestingly, one compound showed dissociation rates from the ternary complex with RT mutants K103N and Y181I 10-20-fold slower than from the corresponding complex with wild-type RT.
authors.affiliation: Istituto di Genetica Molecolare IGM-CNR via Abbiategrasso 207, 27100 Pavia, Italy.
authors.forename: Alberta
authors.initials: A
authors.lastname: Samuele
authors.forename: Marcella
authors.initials: M
authors.lastname: Facchini
authors.forename: Dante
authors.initials: D
authors.lastname: Rotili
authors.forename: Antonello
authors.initials: A
authors.lastname: Mai
authors.forename: Marino
authors.initials: M
authors.lastname: Artico
authors.forename: Mercedes
authors.initials: M
authors.lastname: Armand-Ugón
authors.forename: José A
authors.initials: JA
authors.lastname: Esté
authors.forename: Giovanni
authors.initials: G
authors.lastname: Maga
mesh_terms: D019380:Anti-HIV Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D001665:Binding Sites / Q000187:drug effects / Q000235:genetics; D001667:Binding, Competitive / Q000187:drug effects; D002384:Catalysis; D004305:Dose-Response Relationship, Drug; D024882:Drug Resistance, Viral; D005464:Fluorobenzenes; D020869:Gene Expression Profiling; D054303:HIV Reverse Transcriptase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics*; D007700:Kinetics; D015394:Molecular Structure; D009154:Mutation; D016133:Polymerase Chain Reaction; D011744:Pyrimidinones / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics; D018894:Reverse Transcriptase Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D013237:Stereoisomerism; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D013997:Time Factors
chemical_list: C506288:6-(1-(2,6-difluorophenyl)ethyl)-2-(dimethylamino)-5-methylpyrimidin-4(3H)-one; D019380:Anti-HIV Agents; D005464:Fluorobenzenes; C533731:MC1838; C533732:MC1839; D011744:Pyrimidinones; D011994:Recombinant Proteins; D018894:Reverse Transcriptase Inhibitors; D054303:HIV Reverse Transcriptase