Significant increase in life-threatening infections caused by Gram-negative ‘superbugs’, rising antibiotic resistance and a dearth of novel antibiotics have developed a ‘perfect-storm'. There is an urgency to call for out-of-the-box thinking and explore novel antibiotics to eradicate resistant bacteria. Finding new natural scaffolds with a unique mechanism of action is a promising approach for developing new anti-infective therapeutics. Recent studies have identified a unique cyclic lipodepsipeptide Laterocidine that is active against multidrug-resistant (MDR) Gram-negative pathogens, especially Pseudomonas aeruginosa.1 We have developed the first-ever total chemical synthesis methodology for Laterocidine, which has enabled us to undertake a systematic structure-activity relationship (SAR) study. We have discovered important structural features that drive the antimicrobial activity of Laterocidine using a chemical biology approach and currently conducting lead optimisation for preclinical in vitro and in vivo evaluations.