Compound synthesis

All compounds used in this study were prepared as described in US patent 10,882,272 granted on 5 January 2021. The syntheses of representative compounds AN14353 and AN15368 are described in Supplementary Text 1.

Parasites and mice

C57BL/6J mice were purchased from the Jackson Laboratory and B6.129S7-Ifngtm1Ts/J (IFN-gamma deficient) were bred in-house at the University of Georgia Animal Facility. The SKH-1 ‘hairless’ mice backcrossed to C57BL/6J were a gift from Dr Lisa DeLouise (University of Rochester). All the animals were maintained at the University of Georgia Animal Facility under specific-pathogen-free conditions at 22 °C, 50% humidity and in a 12:12 h light:dark cycle. Male and female mice 6–9 weeks of age were used. All mouse experiments were carried out in strict accordance with the Public Health Service Policy on Humane Care and Use of Laboratory Animals and the Association for Assessment and Accreditation of Laboratory Animal Care accreditation guidelines. The protocol was approved by the University of Georgia Institutional Animal Care and Use Committee (IACUC). T. cruzi tissue culture trypomastigotes of the wild-type Brazil strain, Colombiana strain co-expressing firefly luciferase and tdTomato reporter proteins⁴, and CL strain expressing the fluorescent protein tdTomato¹¹ were maintained through passage in Vero cells (American Type Culture Collection) cultured in RPMI 1640 medium with 10% fetal bovine serum at 37 °C in an…