Lipid droplets (LDs) are the universal lipid storage organelles [1, 2]. Lipids remobilized from LDs are used both for energy production via beta-oxidation or anabolic reactions, such as membrane biosynthesis [–5]. We have previously described three probes ML206, ML219, and ML220, which were optimized from high-throughput screening hits of Drosophila melanogaster embryonic cells and showed a potent LD reduction phenotype, with EC50s of 8 nM, 2 nM, and 705 nM, respectively. Herein, we describe the further optimization of ML219 and characterization of the new probe, ML360. This compound showed potent inhibition of LD formation in the primary Drosophila melanogaster cell line as well as three mammalian cell lines (3T3-L1, COS-7, and AML12). It retained potent activity in 3T3-L1 pre-adipocyte to adipocyte differentiation conditions and showed no signs of cytotoxicity over this extended protocol.