Hunting for PaNACEAs

Chris WatersDepartment of Microbiology and Molecular Genetics

What would be the ideal activity for an anti-biofilm compound?

+ Compound X

Potent and Novel Anti-biofilm Compounds that Enhance Antibiotics (PaNACEAs)

Enhance tobramycin killing of P. aeruginosa biofilms

Tobramycin

• Aminoglycoside: binds to bacterial 30S ribosomal subunit impairing protein synthesis.

• Ototoxicity and nephrotoxicityCystic fibrosis

384 Pin tool + 384-well growth plate

Rinse plate

Treatment plate +250 mg/mL tobramycin (planktonic MIC-1 mg/mL)

Rinse plate

The PaNACEA Screening Protocol

BacTiterGlo

Summary of the pilot screen

Performed pilot screen of 6,080 biologically active compounds at the Center for Chemical Genomics.

Z-factor 0.6 (very good for actual biofilms!)

TRICLOSAN

• Colgate Total= 0.3% = 10 mM

• Mechanism of action: inhibits fatty acid synthesisfabI

• P. aeruginosa known to be resistant

CF11 CF13 CF78 CF99 CF101 CF144 CF1530

20

40

60

80

100

120

TriTobTri/Tob

P. aeruginosa CF Strain

Perc

ent B

iofil

m K

illin

g

Triclosan/Tobramycin effectively kills the biofilms of 19/19 clinical isolates tested

Before treatment

Tobramycin Triclosan Water To-bramycin+Tr

iclosan

P. aerugi-nosa

250000000 7611250 4928200 7391350 3860

3.2E+03

3.2E+04

3.2E+05

3.2E+06

3.2E+07

3.2E+08

CFU/mouse

Triclosan/Tobramycin effective in vivo

Future Directions

1. Screen the additional compounds libraries at the CCG

2. Identify mechanism of triclosan

3. Further characterize additional PaNACEAs

4. Move triclosan/tobramycin treatment forward

Waters LabAlessandra Hunt, PhDWill Soto, PhD

Eric BrugerNico FernandezMichael MaidenBen PursleyGeoff SeverinMeng ShiehRudy Sloup

Jake GibsonCharnay GlossJenny NybergJohn Shook

Alumni: Lauren Priniski!

CCG-University of MichiganMartha LarsenTom McQuadeDavid ShermanPamela SchultzCarl AverangAvi Raveh

Princeton UniversityMartin SemmelhakWei-wei LaoJiaqiang DongEric Kim