Project Summary/Abstract A major challenge facing the future of human health is the rise of antibiotic-resistant pathogenic bacteria. In 2017 the World Health Organization (WHO) published a list of twelve pathogens that will pose the greatest threat to human health due to resistance to frontline treatments. The overall goal of this project will be to identify novel fungal natural products that exhibit antibiotic activity towards the WHO pathogens. Natural products are secondary metabolites produced by living organisms. These small molecule chemical entities have played an important role in traditional medicine for thousands of years and are an essential part of the current therapeutic arsenal for modern medicine. This project will achieve the overall goal through two areas of research. The first area involves the development of a rapid dereplication method that reduces redundant fungal strains in culture libraries to streamline biological screening. The second involves the identification of antibiotic drug-leads with novel chemical scaffolds or that function through a unique antivirulence mechanism of action against the WHO pathogens. Once completed, these two areas of research will be bridged to develop a fungal metabolite drug discovery platform that will be used to promote future lines of biomedical research.