Plasmodium falciparum dihydroorotate dehydrogenase (PfDHODH) has been clinically validated as a target for antimalarial drug discovery, as a triazolopyrimidine class inhibitor (DSM265) is currently undergoing clinical development. Here, we have identified new hydroxyazole scaffold-based PfDHODH inhibitors belonging to two different chemical series. The first series was designed by a scaffold hopping strategy that exploits the use of hydroxylated azoles. Within this series, the hydroxythiadiazole 3 was identified as the best selective PfDHODH inhibitor (IC50 12.0 μM). The second series was designed by modulating four different positions of the hydroxypyrazole scaffold. In particular, hydroxypyrazoles 7e and 7f were shown to be active in the low μM range (IC50 2.8 and 5.3 μM, respectively). All three compounds, 3, 7e and 7f showed clear selectivity over human DHODH (IC50 > 200 μM), low cytotoxicity, and retained micromolar activity in P. falciparum-infected erythrocytes. The crystallographic structures of PfDHODH in complex with compounds 3 and 7e proved their binding mode, supplying essential data for future optimization of these scaffolds. Display omitted •Plasmodium falciparum DHODH is a validated target for malaria treatment.•New PfDHODH inhibitors are designed and synthetized using hydroxyazole scaffolds.•X-ray crystallography determined the binding mode inside the PfDHODH binding site.•Selectively over the human isoform, the hydroxypyrazole 7e inhibit PfDHODH (IC50 = 2.8 μM).•3 best compounds were tested against Pf-infected erythrocytes (EC50 between 26 and 40 μM).