UW-L

1. UW-L MYCOPHYTE DISCOVERY GROUP Chemistry Microbiology Biology Fighting Infection in Patients with Cystic Fibrosis Bill Schwan UW-La Crosse University of Wisconsin–La Crosse

2. Chemistry Microbiology Biology UW-L MYCO-PHYTE DISCOVERY GROUP a UW-La Crosse interdisciplinary initiative Chemistry Microbiology Biology University of Wisconsin–La Crosse

3. RELATIVELY RAPID (and inexpensive) MUCH SLOWER (and costly) UW-L MYCOPHYTE DISCOVERY GROUP General procedure: collect  screen  isolate  screen  elucidate  synthesize  derivatize screen  screen  screen Chemistry Microbiology Biology X inactive–stop X inactive–stop active partition & bioassay crude extract Isolate active cmpd(s) purify solve structure search literature protect IP myco / phyto active–go initial bioassay (other screens?) inactive–stop more bio (and tox) screens synthesis analogs protect IP pre-clinical studies clinical trials University of Wisconsin–La Crosse

4. Prof Tom Volk Biology, UW-L Prof Marc Rott Microbiology, UW-L Prof William Schwan Microbiology, UW-L Prof Aaron Monte Chemistry, UW-L Dr. Joe Toce Chemistry, UW-L Prof James Cook & M. S. Kabir Chemistry, UW-M Dr. Xueting Liu Chemistry, UW-L The WiSys Team: Mindy, Bill Maliyakal, Bethany, Lisa Jonathan Fritz, JD, MS, BS Whyte, Hirschboeck, & Dudek, S.C. MYCOPHYTE DISCOVERY UW-L the multidisciplinary team—these people and more! Chemistry Microbiology Biology University of Wisconsin–La Crosse

5. UW-L MYCOPHYTE DISCOVERY Chemistry Microbiology Biology progress / collaborative opportunities NOVEL ANALOGS PROJECT ACTIVITY STRUCTURE NOVEL? SYNTHESIS PRE-CLINICAL CLINICAL * + ~300 wild mushroom species with wide variety of activities… + ongoing collaborations w/ WI Native Americans / herbal remedies… COULD SHARE CRUDE EXTRACT LIBRARY TO SCREEN FOR ADDL ACTIVITIES NEED HELP HERE AND HERE! University of Wisconsin–La Crosse

6. UW-L MYCOPHYTE DISCOVERY GROUP Chemistry Microbiology Biology Project MD-2/F250 Screened fungal fruiting body library against bacteria that colonize the lungs of Cystic Fibrosis patients Goal: Find new antibiotic University of Wisconsin–La Crosse

7. UW-L MYCOPHYTE DISCOVERY GROUP Chemistry Microbiology Biology What is Cystic Fibrosis? www.nlm.nih.gov University of Wisconsin–La Crosse

8. UW-L MYCOPHYTE DISCOVERY GROUP Chemistry Microbiology Biology What is Cystic Fibrosis? • Homozygous recessive genetic disease • afflicting Caucasian individuals • Average lifespan of a CF patient is only 32 years • Characterized by chronic bronchial infections that • kill more than 90% of CF patients • Antibiotic resistance in the bacteria a big problem University of Wisconsin–La Crosse

9. UW-L MYCOPHYTE DISCOVERY GROUP Chemistry Microbiology Biology Screening Protocol • Screened a library of 260 fungal fruiting bodies • Used disk diffusion assays for screening • Target organisms were Pseudomonas aeruginosa • and Acinetobacter baumannii • Best hit was against fungus labeled F10/F250 • Expanded the assay to include • Burkholderia cepacia and • Stenotrophomonas maltophilia University of Wisconsin–La Crosse

10. UW-L MYCOPHYTE DISCOVERY GROUP Chemistry Microbiology Biology Disk Diffusion Assays Against Glucose Nonfermenting Gram-negative Rods University of Wisconsin–La Crosse

11. UW-L MYCOPHYTE DISCOVERY GROUP Chemistry Microbiology Biology Purification of the Active Drug • First, thin-layer chromatography • Second, HPLC separation of active peak • Elucidation of structure of active drug using • Mass Spectrometry • NMR Spectroscopy University of Wisconsin–La Crosse

12. UW-L MYCOPHYTE DISCOVERY GROUP Chemistry Microbiology Biology Structure of Active Natural Product Drug University of Wisconsin–La Crosse

13. UW-L MYCOPHYTE DISCOVERY GROUP Chemistry Microbiology Biology 2-AQ MICs Against Glucose Nonfermenting Gram-negative Rods University of Wisconsin–La Crosse

14. UW-L MYCOPHYTE DISCOVERY GROUP Chemistry Microbiology Biology What To Do Next? • Synthesize 2-AQ • Make analogs of 2-AQ • Proceed with pre-clinical testing of 2-AQ • More cytotoxicity testing in vitro • Safety testing in mice/rats • Efficacy testing in mice/rats University of Wisconsin–La Crosse

15. UW-L MYCOPHYTE DISCOVERY GROUP Chemistry Microbiology Biology Possible 2-AQ Analogs University of Wisconsin–La Crosse

16. UW-L MYCOPHYTE DISCOVERY GROUP Chemistry Microbiology Biology Synthetic Route #1 Merck Patent Approach University of Wisconsin–La Crosse

17. UW-L MYCOPHYTE DISCOVERY GROUP Chemistry Microbiology Biology Synthetic Route #2 Wittig-Style Approach University of Wisconsin–La Crosse

18. UW-L MYCOPHYTE DISCOVERY GROUP Chemistry Microbiology Biology Acknowledgements Mycophyte Discovery Co-Founders: Aaron Monte, Marc Rott, Joe Toce, Tom Volk UW-L Helpers: Craig Dunek, Kathleen Engelbrecht, Mike Gebhardt, Aaron McCoy, Ron McKelvey, Maria Drieta, Will Fiers, Nathan Connell Others: David Lewis, Jim Cook, Maliyakal John Monetary Assistance: UW-L, UW-System ARG and ARG-WiTAG grants University of Wisconsin–La Crosse

19. become a player in WI biotech? be the first startup in Western WI? …can it be done? UW-L MYCOPHYTE DISCOVERY business opportunities? Chemistry Microbiology Biology University of Wisconsin–La Crosse