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UW-LChemistry
Microbiology
Biology
University of Wisconsin–La Crosse
MYCOPHYTE DISCOVERY GROUP
Fighting Infection in Patients with Cystic Fibrosis
Bill SchwanUW-La Crosse
UW-LChemistry
Microbiology
Biology
MYCO-PHYTE DISCOVERY GROUPa UW-La Crosse interdisciplinary initiative
Chemistry
Microbiology
Biology
University of Wisconsin–La Crosse
UW-LChemistry
Microbiology
Biology
University of Wisconsin–La Crosse
General procedure: collect screen isolate screen elucidate synthesize derivatize screen screen screen
myco / phyto
crudeextract
X inactive–stop
X inactive–stop
active
partition&
bioassay
initial bioassay(other
screens?)
RELATIVELY RAPID(and inexpensive)
MUCH SLOWER(and costly)
more bio (and tox)screens
synthesis analogs protect IP pre-clinical studies clinical trials
MYCOPHYTE DISCOVERY GROUP
active–go
inactive–stop
Isolate active cmpd(s) purify solve structure search literature protect IP
Structure Chemist
Pharmacologist
Patent Lawyers
Chemist
Ethnobotanist
Pharmacologist Classical Botanist
Field Botanist
Medicinal Chemist
$$$$
1. Folk history
2. Tentative identity
3. Initial blind screen forpharmacological activity
4. Confirm identity(Genus, species)
5. Extraction of active molecules
6. Classic pharmacological evaluation (-unique activity?)
7. Structure determination(-unique molecule?)
8. Structure modification(new, better drugs?)
9. Document clinical effectiveness10. Patent synthesis / plant material?$
UW-LChemistry
Microbiology
Biology
MYCOPHYTE DISCOVERYthe multidisciplinary team—these people and more!
Prof William SchwanMicrobiology, UW-L
Prof Tom VolkBiology, UW-L
Prof Marc RottMicrobiology, UW-L
Prof Aaron MonteChemistry, UW-L
The WiSys Team: Mindy, BillMaliyakal, Bethany, Lisa
Jonathan Fritz, JD, MS, BSWhyte, Hirschboeck, & Dudek, S.C.
Dr. Joe ToceChemistry, UW-L
Prof James Cook & M. S. KabirChemistry, UW-MDr. Xueting Liu
Chemistry, UW-L
University of Wisconsin–La Crosse
UW-LChemistry
Microbiology
Biology
University of Wisconsin–La Crosse
progress / collaborative opportunities
MYCOPHYTE DISCOVERY
MD-1Gm+, VRE, MRSA, TB
solved: 2 active compounds
no yes yes underway —
MD-2 Gm– rods solved: 2-AQ no underway no underway —
MD-3Gm+, potent
VRE, MRSApartial solution:
complexprobably — — — —
MD-4 TB solved: grifolin no — — — —
MD-5potent, broad
antifungal, etcpurification underway
— — — — —
MD-6 broad, potentpurification underway
— — — — —
MD-7N. sicca / gonorrhoeae
purification underway
— — — — —
— — — — — —
— — — — — —
PROJECT ACTIVITY STRUCTURE NOVEL? SYNTHESISNOVELANALOGS
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
AND HERE!NEED HELP HERE
*
UW-LChemistry
Microbiology
Biology
University of Wisconsin–La Crosse
MYCOPHYTE DISCOVERY GROUP
Screened fungal fruiting body library against bacteria that colonize the lungs of Cystic Fibrosis patients
Goal: Find new antibiotic
Project MD-2/F250
UW-LChemistry
Microbiology
Biology
University of Wisconsin–La Crosse
MYCOPHYTE DISCOVERY GROUP
www.nlm.nih.gov
What is Cystic Fibrosis?
UW-LChemistry
Microbiology
Biology
University of Wisconsin–La Crosse
MYCOPHYTE DISCOVERY GROUP
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
UW-LChemistry
Microbiology
Biology
University of Wisconsin–La Crosse
MYCOPHYTE DISCOVERY GROUP
• 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
Screening Protocol
UW-LChemistry
Microbiology
Biology
University of Wisconsin–La Crosse
MYCOPHYTE DISCOVERY GROUP
Disk Diffusion Assays Against Glucose Nonfermenting Gram-negative Rods
UW-LChemistry
Microbiology
Biology
University of Wisconsin–La Crosse
MYCOPHYTE DISCOVERY GROUP
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
UW-LChemistry
Microbiology
Biology
University of Wisconsin–La Crosse
MYCOPHYTE DISCOVERY GROUP
Structure of Active Natural Product Drug
UW-LChemistry
Microbiology
Biology
University of Wisconsin–La Crosse
MYCOPHYTE DISCOVERY GROUP
2-AQ MICs Against Glucose Nonfermenting
Gram-negative Rods
UW-LChemistry
Microbiology
Biology
University of Wisconsin–La Crosse
MYCOPHYTE DISCOVERY GROUP
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
UW-LChemistry
Microbiology
Biology
University of Wisconsin–La Crosse
MYCOPHYTE DISCOVERY GROUP
Possible 2-AQ Analogs
UW-LChemistry
Microbiology
Biology
University of Wisconsin–La Crosse
MYCOPHYTE DISCOVERY GROUP
Synthetic Route #1
Merck Patent Approach
UW-LChemistry
Microbiology
Biology
University of Wisconsin–La Crosse
MYCOPHYTE DISCOVERY GROUP
Synthetic Route #2
Wittig-Style Approach
UW-LChemistry
Microbiology
Biology
University of Wisconsin–La Crosse
MYCOPHYTE DISCOVERY GROUP
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
MYCOPHYTE DISCOVERY
become a player in WI biotech?
be the first startup in Western WI?
…can it be done?
UW-LChemistry
Microbiology
Biology
University of Wisconsin–La Crosse
business opportunities?
