Summary Malaria remains a major global health problem, causing tremendous morbidity and mortality. Disease occurs during the blood-stage of human infections with asexual proliferation of Plasmodium parasites in red blood cells (RBCs) leading to their destruction. Due to the evolving threat of drug resistance, there is an urgent need to develop new antimalarials for control and eradication efforts. We hypothesize that host-directed inhibitors (HDIs) will circumvent drug resistance in the parasite, which would have to significantly alter its biology to bypass its requirement for the essential host target. An increasing number of studies have reported RBC-specific inhibitors, but these are poorly validated. We propose to identify and validate HDIs. We will include compounds identified by screening a large drug repurposing library with putative targets in the RBC proteome. To validate host targeting as the antimalarial mechanism of action, we will perform synthetic lethal assays using in vitro cultured RBCs (cRBCs) with knockdown of putative targets. Cellular Thermal Shift Assays will be used to demonstrate direct binding to host targets. Finally, for compounds that appear to be bona fide HDIs, we will assess the ability of the parasite to develop resistance. The establishment of a pipeline to validate host targeting as the mechanism of action of putative HDIs will be a significant advance to the field, supporting the development of host-directed therapeutics in the future.