title: A screen against Leishmania intracellular amastigotes: comparison to a promastigote screen and identification of a host cell-specific hit.
abstract: The ability to screen compounds in a high-throughput manner is essential in the process of small molecule drug discovery. Critical to the success of screening strategies is the proper design of the assay, often implying a compromise between ease/speed and a biologically relevant setting. Leishmaniasis is a major neglected disease with limited therapeutic options. In order to streamline efforts for the design of productive drug screens against Leishmania, we compared the efficiency of two screening methods, one targeting the free living and easily cultured promastigote (insect-infective) stage, the other targeting the clinically relevant but more difficult to culture intra-macrophage amastigote (mammal-infective) stage. Screening of a 909-member library of bioactive compounds against Leishmania donovani revealed 59 hits in the promastigote primary screen and 27 in the intracellular amastigote screen, with 26 hits shared by both screens. This suggested that screening against the promastigote stage, although more suitable for automation, fails to identify all active compounds and leads to numerous false positive hits. Of particular interest was the identification of one compound specific to the infective amastigote stage of the parasite. This compound affects intracellular but not axenic parasites, suggesting a host cell-dependent mechanism of action, opening new avenues for anti-leishmanial chemotherapy.
authors.affiliation: Department of Pathology, Sandler Center for Drug Discovery, University of California San Francisco, San Francisco, California, United States of America. Geraldine.DeMuylder@ucsf.edu
authors.forename: Geraldine
authors.initials: G
authors.lastname: De Muylder
authors.forename: Kenny K H
authors.initials: KK
authors.lastname: Ang
authors.forename: Steven
authors.initials: S
authors.lastname: Chen
authors.forename: Michelle R
authors.initials: MR
authors.lastname: Arkin
authors.forename: Juan C
authors.initials: JC
authors.lastname: Engel
authors.forename: James H
authors.initials: JH
authors.lastname: McKerrow
mesh_terms: D000818:Animals; D000981:Antiprotozoal Agents / Q000737:chemistry / Q000494:pharmacology*; D045744:Cell Line, Tumor; D055808:Drug Discovery; D057166:High-Throughput Screening Assays; D006790:Host-Parasite Interactions / Q000187:drug effects; D006801:Humans; D007891:Leishmania / Q000187:drug effects* / Q000254:growth & development; D007896:Leishmaniasis / Q000469:parasitology*; D008018:Life Cycle Stages / Q000187:drug effects; D008264:Macrophages / Q000469:parasitology; D009270:Naloxone / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology
chemical_list: D000981:Antiprotozoal Agents; D054852:Small Molecule Libraries; D009270:Naloxone; C034643:naloxonazine