With emerging resistance to frontline treatments, it is vital that new antimalarial drugs are identified to target Plasmodium falciparum. A critical process during the parasite’s asexual lifecycle is invasion of red blood cells (RBCs), a mechanism that requires many unique parasite proteins that could be exploited as druggable targets. We have recently reported the identification of a compound, MMV020291, as a specific inhibitor of RBC invasion, and successful drug resistance selection was performed. Whole genome sequencing on three MMV020291 resistant populations revealed three non-synonymous single nucleotide polymorphisms (SNPs) in two genes; two populations had SNPs in a gene encoding profilin (N154Y, K124N) and the third had a SNP present in the actin-1 gene (M356L). Using CRISPR-Cas9, we engineered these SNPs into wildtype parasites, reproducing parasite resistance to MMV020291. Monomeric actin-1 is polymerised to form filamentous strands, which are crucial for generating the force required for RBC invasion, with profilin acting to regulate this process by binding monomeric actin-1. MMV020291 appears to inhibit actin polymerisation and we are seeking to understand how the compound engages its target proteins. We have developed a series of MMV020291 analogues, achieving potency with an EC50 <100 nM, indicating the possible development for invasion blocking drugs. In addition, MMV020291 may be utilised a research tool to study the complex actin dynamics in the invading malaria parasite.