The present status of antibiotic research requires an urgent invention of novel agents that act on multidrug resistant bacteria. The World Health Organization (WHO) has classified antibiotic-resistant priority bacterium into critical, high and medium priority according to the urgency of need for new antibiotics. Naturally occurring uridine-derived 'nucleoside antibiotics' have shown promising activity against numerous priority resistant organisms via inhibition of the transmembrane protein MraY (translocase I), which is yet to be explored in a clinical context. The catalytic activity of MraY is an essential process for bacterial cell viability and growth including that of priority organisms. Muraymycins are one subclass of naturally occurring MraY inhibitors. Despite having potent antibiotic properties, the structural complexity of muraymycins advocates for simplified analogues as potential lead structures. Herein we report the systematic structure-activity relationship (SAR) study of serine template linked simplified muraymycin-type analogues. This detailed SAR lead study of serine template analogues has successfully revealed that the complex structure of naturally occurring muraymycins could be easily simplified to afford bioactive scaffolds against resistant priority organisms. This study will pave the way for the development of novel antibacterial lead compounds based on a simplified serine template.