title: Expression, purification, characterization, and in vivo targeting of trypanosome CTP synthetase for treatment of African sleeping sickness.
abstract: African sleeping sickness is a fatal disease caused by two parasite subspecies: Trypanosoma brucei gambiense and T. b. rhodesiense. We previously reported that trypanosomes have extraordinary low CTP pools compared with mammalian cells. Trypanosomes also lack salvage of cytidine/cytosine making the parasite CTP synthetase a potential target for treatment of the disease. In this study, we have expressed and purified recombinant T. brucei CTP synthetase. The enzyme has a higher K(m) value for UTP than the mammalian CTP synthetase, which in combination with a lower UTP pool may account for the low CTP pool in trypanosomes. The activity of the trypanosome CTP synthetase is irreversibly inhibited by the glutamine analogue acivicin, a drug extensively tested as an antitumor agent. There is a rapid uptake of acivicin in mice both given intraperitoneally and orally by gavage. Daily injection of acivicin in trypanosome-infected mice suppressed the infection up to one month without any significant loss of weight. Experiments with cultured bloodstream T. brucei showed that acivicin is trypanocidal if present at 1 mum concentration for at least 4 days. Therefore, acivicin may qualify as a drug with "desirable" properties, i.e. cure within 7 days, according to the current Target Product Profiles of WHO and DNDi.
authors.affiliation: Department of Medical Biochemistry and Biophysics, Umeå University, SE-901 87 Umeå, Sweden. artur.fijolek@medchem.umu.se
authors.forename: Artur
authors.initials: A
authors.lastname: Fijolek
authors.forename: Anders
authors.initials: A
authors.lastname: Hofer
authors.forename: Lars
authors.initials: L
authors.lastname: Thelander
mesh_terms: D000818:Animals; D019731:Carbon-Nitrogen Ligases / Q000096:biosynthesis* / Q000737:chemistry*; D003562:Cytidine / Q000737:chemistry; D003596:Cytosine / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D014344:Trypanocidal Agents / Q000494:pharmacology; D014346:Trypanosoma brucei brucei / Q000201:enzymology*; D014347:Trypanosoma brucei gambiense / Q000201:enzymology*; D014353:Trypanosomiasis, African / Q000628:therapy*
chemical_list: D004791:Enzyme Inhibitors; D014344:Trypanocidal Agents; D003562:Cytidine; D003596:Cytosine; D019731:Carbon-Nitrogen Ligases; C019951:CTP synthetase