Anti-Infection Discovery Platform

1. Anti-Infection Discovery Platform New Platform to Combat Bacterial, Fungal, and Parasitic infections Inventors: Jochen Buck, MD, PhD Lonny Levin, PhD Professor of Pharmacology Associate Professor of Pharmacology Professors Buck and Levin discovered mammalian soluble adenylyl cyclase as the nature’s CO2 sensor. They collaborate and pool their NIH grants to jointly run a 15-member research lab.

2. Anti-Infection Discovery Platform Market Bacterial infection ~$20B Fungal infection ~$2B Malaria ~$500M Applications may encompass both human and veterinarian space.

3. Anti-Infection Discovery Platform Prior to 1999 Adenylyl cyclase (AC) generates cyclic AMP – ubiquitous second messenger system conserved from bacteria to man Mammals had transmembrane AC simpler life forms have soluble AC as well In 1999 Cornell inventors discovered mammalian soluble AC (sAC), and found that it is CO2 responsive – functions as a key metabolic sensor. More recently, inventors studied CO2 responsiveness of sAC from microbes – found same paradigm

4. Air 5% CO2 Anti-Infection Discovery Platform CO2 concentration is more than 150-fold higher (5%) inside mammals than in air (0.033%). Pathogenic microbes use CO2-sensing sAC to determine if they are in air or in a host. In a host, pathogenic processes are set off, including morphological changes, advances through life cycle, and toxin secretion. E. Coli. Lane 3 is 5% CO2; P80 is Tir, a secreted toxin Plasmodium maturation in red blood cells requires high CO2 C. albicansfilamentation

5. Normal bugs Pathogenic bugs All bugs die Drug Treatment Pathogenic Process Anti-Infection Discovery Platform Current Anti-infection Paradigm New Paradigm Normal bugs Pathogenic bugs Normal bugs remain alive. Pathogenic Process Drug Treatment Inhibiting CO2-sensing sAC generally blocks pathogenesis but not necessarily killing the microbes. Does not harm normal flora and may decrease tendency for resistance to evolve.

6. Anti-Infection Discovery Platform Technology Status • Validated concept in Candida albicanswith knockout • Cloned relevant CO2-sensing sAC from C. albicans, an opportunistic pathogen in human,and used for screening • Cloned relevant CO2-sensing sAC fromPseudomonas aeruginosa, an opportunistic human and plant pathogen, and used for screening • Established HTS screen for sAC inhibitors, with counterscreen against human sAC • Identified nonproprietary, microbe-selective hits (3rd generation library) • Tested hits against Plasmodium falciparum, an obligatory parasite that causes malaria, cultivated in human red blood cells – killed them • Tested hits against live Trypanosoma brucei, an obligatory protist that causes sleeping disease– prevented growth • Cloning of CO2-sensing sAC from other pathogens for HTS underway at Cornell

7. Anti-Infection Discovery Platform • Intellectual Property • Issued US Patent on mammalian sAC for counterscreen (US6544768) • Applications pending in US, EP, China, and India for malaria and fungal infection (nationalized PCT WO2005070419 – note published apps do not reflect preliminary amendments) • Provisional applications filed for broad uses of platform • Publications • Mogensen EG, et al. (2006) Cryptococcus neoformans senses CO2 through the carbonic anhydrase Can2 and the adenylyl cyclase Cac1. Eukaryot Cell. 5(1):103-11. • Klengel T, et al (2005) Fungal adenylyl cyclase integrates CO2 sensing with cAMP signaling and virulence. Curr Biol. 15(22):2021-6. • Bank et al, (Submitted) Killing malaria parasites by inhibiting the Plasmodium falciparum carbon dioxide sensor.