title: Configurations of diastereomeric hydroxyethylene isosteres strongly affect biological activities of a series of specific inhibitors of human-immunodeficiency-virus proteinase.
abstract: Human immunodeficiency virus (HIV) proteinase (PR) represents an important target for antiviral chemotherapy. We present an analysis of inhibitory activities of a series of pseudopeptide inhibitors of HIV-1 PR. All inhibitors were N-protected tetrapeptides with the scissile bond replaced by a nonhydrolysable hydroxyethylene or hydroxyethylamine isostere. To elucidate subtle structural requirements of the PR binding cleft, we synthesised inhibitors with four combinations of configurations at the asymmetric carbons of the isostere. Compounds were tested in vitro using purified recombinant enzyme and a chromogenic peptide substrate. The differences in inhibition constants between individual diastereoisomers reached three orders of magnitude. The most active hydroxyethylene-containing inhibitor possessed the 2R,4S,5S configuration at the isostere. Inhibitor activity was also tested in mammalian cell culture by analysing reduction of viral polyprotein processing and virus infectivity. The results obtained in tissue culture were generally in agreement with the in vitro data, giving a similar order of potency for the individual diastereoisomers. The most active compounds completely blocked production of infectious virus. A simulation method for interaction was employed to build a model of the inhibitors in the PR active site, to identify the interactions responsible for the differences in activities of individual stereoisomers, and to estimate the relative contribution of individual structural features to the overall inhibitory activity.
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Science of the Czech Republic, Praha. konval@uochb.cas.cz
authors.forename: J
authors.initials: J
authors.lastname: Konvalinka
authors.forename: J
authors.initials: J
authors.lastname: Litera
authors.forename: J
authors.initials: J
authors.lastname: Weber
authors.forename: J
authors.initials: J
authors.lastname: Vondrásek
authors.forename: M
authors.initials: M
authors.lastname: Hradílek
authors.forename: M
authors.initials: M
authors.lastname: Soucek
authors.forename: I
authors.initials: I
authors.lastname: Pichová
authors.forename: P
authors.initials: P
authors.lastname: Majer
authors.forename: P
authors.initials: P
authors.lastname: Strop
authors.forename: J
authors.initials: J
authors.lastname: Sedlácek
authors.forename: A M
authors.initials: AM
authors.lastname: Heuser
authors.forename: H
authors.initials: H
authors.lastname: Kottler
authors.forename: H G
authors.initials: HG
authors.lastname: Kräusslich
mesh_terms: D000818:Animals; D001665:Binding Sites; D019556:COS Cells; D017320:HIV Protease Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D015497:HIV-1 / Q000187:drug effects*; D008958:Models, Molecular; D013237:Stereoisomerism; D013329:Structure-Activity Relationship
chemical_list: D017320:HIV Protease Inhibitors