Malaria is a devastating disease caused by plasmodium parasite carried by mosquitoes. Approximately 40% of the world’s population lives in the area at the risk of malaria and it is one of the main causes of morbidity and mortality in many developing countries (Central-south America, many parts of Asia, and Africa). Malaria is recognized as one of the reasons of poverty in developing countries and infection caused by malaria reduced their economic growth by 1.3% per year. Currently, number of synthetic and semisynthetic drugs from Quinoline, Antifolate, Artemisinin groups as well as number of antibiotics and Atovaquone are used in clinics for the treatment of malaria.However, startling spread of drug-resistant malaria to practically all currently available drugs have been observed. Resistance in regions threatened by malaria continues to spread, indicating current therapeutic agents will be practically ineffective in the near future. This has reinforced the search for new drugs with a novel mode of action and PfSUB1 inhibition could be targeted to overcome the resistance with established chemotherapies. Plasmodium falciparum subtilisin-like serine protease 1 (PfSUB1) belongs to the subtilisin-like family of serine proteases that leads to rupture of the vacuole and host cell membrane, causing merozoites egress. Therefore PfSUB1 appears to play a critical role in the life cycle of the parasite and hence an attractive target for the development of novel antimalarial drugs. However, the development of small molecule inhibitors for PfSUB1 has turned out to be a difficult task. Screening of a collection of >170,000 low molecular weight compounds resulted in the identification of a structurally complex natural product MRT 12113, quinolylhydrazone derivative, chloroisocoumarin derivative JCP104, difluorostatone based micromolar level PfSUB1 inhibitors.At the time we started the search for PfSUB1 inhibitors only compounds MRT 12113 and JCP104 were reported in literature.