S1 SUPPORTING INFORMATION 3-Aminoquinazolinediones as a New Class of Antibacterial Agents Demonstrating Excellent Antibacterial Activity Against Wild-type and Multi-drug Resistant Organisms. Edmund L. Ellsworth, *, # Tuan P. Tran, # H. D. Hollis Showalter, # Joseph P. Sanchez, # Brian M. Watson, # Michael A. Stier, # John M. Domagala, # Stephen J. Gracheck, # E. Themis Joannides, # Martin A. Shapiro, # Steve A. Dunham, # Debra L. Hanna, # Michael D. Huband, # Jeffrey W. Gage, # Joel C. Bronstein, # Jia Yeu Liu, # Dai Q. Nguyen £ and Rajeshwar Singh £ # Pfizer Global Research and Development, Michigan Laboratories, 2800 Plymouth Rd., Ann Arbor, Michigan 48105, USA and £ NAEJA Pharmaceuticals, 4290-091, Edmonton, Canada T6E5V2 Table of Contents 1.1 Bacterial Cultures 1.2 Permanent Stock Culture Collection 1.3 Maintenance of Stock Cultures 1.4 Confirming Identity of Cultures Standardized Organism Inocula 1.5 Compounds and their Preparation 1.6 Drug Dilution Tray Preparation 1.7 Tray Inoculation 1.8 MIC Test Results 1.9 Drug-Binding/Competition Studies 1.10 Macromolecular Synthesis Inhibition assay 1.11 References Except for clarifying or modifying statements, MIC testing followed procedures recommended by the NCCLS 1-3 or followed the descriptions cited below. 1.1 Bacterial Cultures Bacterial cultures were either recent clinical isolates cultured from patients (primarily in the US) within the past 2 years, in vivo strains, or laboratory strains with specific permeability / resistance characteristics. Incubations were at 35 °C. Stock bacterial cultures were maintained on Tryptic Soy Agar containing 5% Sheep Blood (BD, Becton Dickinson Microbiology Systems, Cockeysville, Maryland) and fastidious organisms were maintained on Chocolate Agar II Plates (BD). Specific conditions of handling are listed below.