title: Rational design of novel antimicrobials: blocking purine salvage in a parasitic protozoan.
abstract: All parasitic protozoa obtain purine nucleotides solely by salvaging purine bases and/or nucleosides from their host. This observation suggests that inhibiting purine salvage may be a good way of killing these organisms. To explore this idea, we attempted to block the purine salvage pathway of the parasitic protozoan Tritrichomonas foetus. T. foetus is a good organism to study because its purine salvage depends primarily on a single enzyme, hypoxanthine-guanine-xanthine phosphoribosyltransferase (HGXPRTase), and could provide a good model for rational drug design through specific enzyme inhibition. Guided by the crystal structure of T. foetus HGXPRTase, we used structure-based drug design to identify several non-purine compounds that inhibited this enzyme without any detectable effect on human HGPRTase. One of these compounds, 4-[N-(3, 4-dichlorophenyl)carbamoyl]phthalic anhydride (referred to as TF1), was selected for further characterization. TF1 was shown to be a competitive inhibitor of T. foetus HGXPRTase with respect to both guanine (in the forward reaction; Ki = 13 microM) and GMP (in the reverse reaction; Ki = 10 microM), but showed no effect on the homologous human enzyme at concentrations of up to 1 mM. TF1 inhibited the in vitro growth of T. foetus with an EC50 of approximately 40 microM. This inhibitory effect was associated with a decrease in the incorporation of exogenous guanine into nucleic acids, and could be reversed by supplementing the growth medium with excess exogenous hypoxanthine or guanine. Thus, rationally targeting an essential enzyme in a parasitic organism has yielded specific enzyme inhibitors capable of suppressing that parasite's growth.
authors.affiliation: Department of Biochemistry and Biophysics, University of California, San Francisco 94143-0448, USA.
authors.forename: J R
authors.initials: JR
authors.lastname: Somoza
authors.forename: A G
authors.initials: AG
authors.lastname: Skillman
authors.forename: N R
authors.initials: NR
authors.lastname: Munagala
authors.forename: C M
authors.initials: CM
authors.lastname: Oshiro
authors.forename: R M
authors.initials: RM
authors.lastname: Knegtel
authors.forename: S
authors.initials: S
authors.lastname: Mpoke
authors.forename: R J
authors.initials: RJ
authors.lastname: Fletterick
authors.forename: I D
authors.initials: ID
authors.lastname: Kuntz
authors.forename: C C
authors.initials: CC
authors.lastname: Wang
mesh_terms: D000818:Animals; D000994:Antitrichomonal Agents / Q000138:chemical synthesis* / Q000494:pharmacology*; D001667:Binding, Competitive / Q000187:drug effects; D018360:Crystallography, X-Ray; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000494:pharmacology; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D008958:Models, Molecular; D011684:Purine Nucleosides / Q000037:antagonists & inhibitors* / Q000378:metabolism; D016835:Tritrichomonas foetus / Q000201:enzymology*
chemical_list: D000994:Antitrichomonal Agents; D004791:Enzyme Inhibitors; D011684:Purine Nucleosides; D007041:Hypoxanthine Phosphoribosyltransferase