biostudies-literatureSakura TScience and Technology Research Partnership for Sustainable DevelopmentJapan Agency for Medical Research and DevelopmentMinistry of Education, Culture, Sports, Science and TechnologyShionogiMinistry of Health and WelfareJapan Society for the Promotion of Science4115-4126https://www.ebi.ac.uk/biostudies/studies/S-EPMC11650643biostudies-literatureUnknown10(12)The urgent need for rapidly acting compounds in the development of antimalarial drugs underscores the significance of such compounds in overcoming resistance issues and improving patient adherence to antimalarial treatments. The present study introduces a high-throughput screening (HTS) approach using 1536-well plates, employing <i>Plasmodium falciparum</i> lactate dehydrogenase (PfLDH) combined with nitroreductase (NTR) and fluorescent probes to evaluate inhibition of the growth of the asexual blood stage of malaria parasites. This method was adapted to efficiently assess the speed of action profiling (SAP) in a 384-well plate format, streamlining the traditionally time-consuming screening process. By successfully screening numerous compounds, this approach identified fast-killing hits early in the screening process, addressing challenges associated with artemisinin-based combination therapies. The high-throughput SAP method is expected to be of value in continuously monitoring fast-killing properties during structure-activity relationship studies, expediting the identification and development of novel, rapidly acting antimalarial drugs within phenotypic drug discovery campaigns.ACS infectious diseasesAccelerating Antimalarial Drug Discovery with a New High-Throughput Screen for Fast-Killing Compounds.PMC11650643JP14425718JP1681136222K07045JP1000028420H00620JP2251204723fk010868021fk010813823H02711Sakura TInaoka DKYoshida EKato TIshii RKita KfalseAccelerating Antimalarial Drug Discovery with a New High-Throughput Screen for Fast-Killing Compounds.The urgent need for rapidly acting compounds in the development of antimalarial drugs underscores the significance of such compounds in overcoming resistance issues and improving patient adherence to antimalarial treatments. The present study introduces a high-throughput screening (HTS) approach using 1536-well plates, employing <i>Plasmodium falciparum</i> lactate dehydrogenase (PfLDH) combined with nitroreductase (NTR) and fluorescent probes to evaluate inhibition of the growth of the asexual blood stage of malaria parasites. This method was adapted to efficiently assess the speed of action profiling (SAP) in a 384-well plate format, streamlining the traditionally time-consuming screening process. By successfully screening numerous compounds, this approach identified fast-killing hits early in the screening process, addressing challenges associated with artemisinin-based combination therapies. The high-throughput SAP method is expected to be of value in continuously monitoring fast-killing properties during structure-activity relationship studies, expediting the identification and development of novel, rapidly acting antimalarial drugs within phenotypic drug discovery campaigns.2024-01-01T00:00:00Z2024 Dec2025-04-03T23:07:01.309Z2025-04-03T23:07:01.309ZS-EPMC116506433956129910.1021/acsinfecdis.4c00328