title: P. falciparum in vitro killing rates allow to discriminate between different antimalarial mode-of-action.
abstract: Chemotherapy is still the cornerstone for malaria control. Developing drugs against Plasmodium parasites and monitoring their efficacy requires methods to accurately determine the parasite killing rate in response to treatment. Commonly used techniques essentially measure metabolic activity as a proxy for parasite viability. However, these approaches are susceptible to artefacts, as viability and metabolism are two parameters that are coupled during the parasite life cycle but can be differentially affected in response to drug actions. Moreover, traditional techniques do not allow to measure the speed-of-action of compounds on parasite viability, which is an essential efficacy determinant. We present here a comprehensive methodology to measure in vitro the direct effect of antimalarial compounds over the parasite viability, which is based on limiting serial dilution of treated parasites and re-growth monitoring. This methodology allows to precisely determine the killing rate of antimalarial compounds, which can be quantified by the parasite reduction ratio and parasite clearance time, which are key mode-of-action parameters. Importantly, we demonstrate that this technique readily permits to determine compound killing activities that might be otherwise missed by traditional, metabolism-based techniques. The analysis of a large set of antimalarial drugs reveals that this viability-based assay allows to discriminate compounds based on their antimalarial mode-of-action. This approach has been adapted to perform medium throughput screening, facilitating the identification of fast-acting antimalarial compounds, which are crucially needed for the control and possibly the eradication of malaria.
authors.affiliation: Tres Cantos Medicine Development Campus, Diseases of the Developing World, GlaxoSmithKline, Tres Cantos, Madrid, Spain.
authors.forename: Laura M
authors.initials: LM
authors.lastname: Sanz
authors.forename: Benigno
authors.initials: B
authors.lastname: Crespo
authors.forename: Cristina
authors.initials: C
authors.lastname: De-Cózar
authors.forename: Xavier C
authors.initials: XC
authors.lastname: Ding
authors.forename: Jose L
authors.initials: JL
authors.lastname: Llergo
authors.forename: Jeremy N
authors.initials: JN
authors.lastname: Burrows
authors.forename: Jose F
authors.initials: JF
authors.lastname: García-Bustos
authors.forename: Francisco-Javier
authors.initials: FJ
authors.lastname: Gamo
mesh_terms: D000818:Animals; D000962:Antimalarials / Q000494:pharmacology*; D018929:Cell Culture Techniques / Q000379:methods; D004353:Drug Evaluation, Preclinical; D006801:Humans; D019271:Hypoxanthine / Q000737:chemistry; D020128:Inhibitory Concentration 50; D016778:Malaria, Falciparum / Q000188:drug therapy*; D010963:Plasmodium falciparum / Q000187:drug effects* / Q000378:metabolism*; D015203:Reproducibility of Results; D013997:Time Factors; D016896:Treatment Outcome
chemical_list: D000962:Antimalarials; D019271:Hypoxanthine