PROJECT ABSTRACT/SUMMARY The viruses belonging to the family Arenaviridae contain several causative agents of hemorrhagic fever in humans. For instance, Lassa virus causes Lassa fever, which is a major public health threat in endemic regions of West Africa. Junin virus, Machupo virus, Guanarito virus, Sabia virus, and Chapare virus cause South American hemorrhagic fevers, severe diseases in humans resulting in high fatality rates. These highly pathogenic arenaviruses are classified as Risk Group-4 agents, which need to be handled in biosafety level-4 facilities due to their high pathogenicity and lack of vaccines or therapeutics. Additionally, arenaviruses causing hemorrhagic fevers are listed on Category A Priority Pathogens because of the highest risk to national security and public health. Thus, countermeasures, such as vaccines or antivirals, against arenavirus infections are crucial medical needs globally. Most antiviral agents are inhibitors of proteases or inhibitors of virus replication. In a search for alternative approaches, we propose to study in more detail the dependence of arenavirus on metabolism of two key monosaccharides, D-glucose and D-mannose. Our focus will be on using carbohydrate mimetics/antimetabolites as antivirals. Specifically, we will employ isotopically labeled monosaccharides and molecular probes that include 2-deoxy-D-glucose (2-DG), a D-glucose, and also a D-mannose analog as a reference compund, and WP1096, a 2-iodo-D-mannose as a novel prototype carbohydrate antimetabolite antiviral. Our results will enhance the success of designing future carbohydrate antimetabolite antiviral compounds as therapeutics to alleviate suffering and contain arenaviral outbreaks. We will address this goal through three specific aims. Specific Aim 1. Determine the effect of WP1096 and 2-DG (as a control) on glycolysis pathways and viral glycoproteins of tissue culture cells infected with arenaviruses (LASV, JUNV and MACV). Specific Aim 2. In vivo evaluation of WP1096 that includes preclinical toxicology, tissue distribution, pharmacokinetics, and analysis of active metabolites. Specific Aim 3. Determine the in vivo protective efficacy of WP1096 and 2-DG (as a control) and analyze their effects on glycosylation patterns and the host glycolysis pathway. Taken together, we hope to develop promising pan-arenavirus therapeutics and gain the insight for development of broad spectrum antivirals effective against multiple viral families.