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We used a phenotypic high-throughput screening (HTS) platform to identify new antimalarials, uncovering three related pyrrole-, indole-, and indoline-based series with a shared \u03b1-azacyclic acetamide core. These compounds showed fast-killing activity on asexual blood-stage\n                  <jats:italic>Plasmodium falciparum<\/jats:italic>\n                  parasites, were not cytotoxic, and disrupted parasite intracellular pH and Na\n                  <jats:sup>+<\/jats:sup>\n                  regulation similarly to cipargamin (KAE609), a clinically advanced inhibitor of the\n                  <jats:italic>P. falciparum<\/jats:italic>\n                  Na\n                  <jats:sup>+<\/jats:sup>\n                  pump (\n                  <jats:italic>Pf<\/jats:italic>\n                  ATP4).\n                  <jats:italic>Pf<\/jats:italic>\n                  ATP4 is localized to the parasite plasma membrane and is essential for maintaining a low cytosolic Na\n                  <jats:sup>+<\/jats:sup>\n                  concentration. Resistance selections on\n                  <jats:italic>P. falciparum<\/jats:italic>\n                  parasites with two \u03b1-azacyclic acetamide analogs identified mutations in\n                  <jats:italic>Pf<\/jats:italic>\n                  ATP4, and cross-resistance was observed across the \u03b1-azacyclic acetamides and KAE609, confirming\n                  <jats:italic>Pf<\/jats:italic>\n                  ATP4 as the target.\n                  <jats:italic>Pf<\/jats:italic>\n                  ATP4 is a well-established antimalarial target, and identification of additional\n                  <jats:italic>Pf<\/jats:italic>\n                  ATP4 inhibitors provides alternative avenues to disrupt its function.\n                <\/jats:p>","DOI":"10.1101\/2025.05.20.655166","type":"posted-content","created":{"date-parts":[[2025,5,20]],"date-time":"2025-05-20T23:45:20Z","timestamp":1747784720000},"source":"Crossref","is-referenced-by-count":0,"title":["Identification of \u03b1-azacyclic acetamide-based inhibitors of\n                  <i>P. falciparum<\/i>\n                  Na\n                  <sup>+<\/sup>\n                  pump (\n                  <i>Pf<\/i>\n                  ATP4) with fast-killing asexual blood-stage antimalarial activity by phenotypic screening"],"prefix":"10.64898","author":[{"suffix":"Jr","given":"Arturo","family":"Casas","sequence":"first","affiliation":[{"name":"Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX, 75390, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leah S.","family":"Imlay","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX, 75390, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vandana","family":"Thathy","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Columbia University Irving Medical Center, New 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