Inhibition of Mycobacterium tuberculosis RecA · responsible for approximately 1.6 million deaths...
Transcript of Inhibition of Mycobacterium tuberculosis RecA · responsible for approximately 1.6 million deaths...
RESEARCH POSTER PRESENTATION DESIGN © 2015
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• Mycobacterium tuberculosis (Mtb) infects more than 10 million new individuals and is responsible for approximately 1.6 million deaths per year globally1.
• Tuberculosis is curable, but drug regimens are long, 6-20 months, and tuberculosis often persists in communities where medical care is largely inaccessible2.
• Drug resistance in Mtb is rising, nullifying the effects of first and second-line antibiotics and creating a need for new drugs3.
• Under stress, persistent bacteria can become more physiologically tolerant to drugs, although no mutations have been acquired, by activating protein signaling pathways that initiate an SOS response that leads to resistant-like phenotypes - giving the bacteria more time to acquire resistance-conferring mutations4.
• Choudhary et. al. found that RecA/LexAexpression is increased in response to fluoroquinolones in Mtb, which leads to the activation of an SOS response and increased drug tolerance5.
• Inhibition of RecA sensitized the persistersto antibiotics.
• Although there is an available inhibitor, suramin, it binds at 50 μM affinity, which is considered too high for use in the clinic.
• The development of high affinity inhibitory ligands against RecA could be used to sensitize Mtb populations tolerant to first-line drugs, extending the usability of these drugs and relieving pressure to come up with newer drugs to overcome antimicrobial resistance.
Inhibition of Mycobacterium tuberculosis RecA
2Atomwise, 717 Market Street, Suite 800, San Francisco, CA 94103
Christopher Beaudoin1*, Terry O’Brien2, Tom Blundell1
ACKNOWLEDGMENTS
Antibiotic Research UK
Marcin Skwark
BACKGROUND METHODS FUTURE• Validate compounds with ATP-binding affinity
functional assays
• Work with Atomwise chemists to potentially synthesize derivatives of the successful compounds with higher affinity
• Test compounds in vivo using Mtb cell culture
180 Tennis Court Road, Department of Biochemistry, University of Cambridge CB2 1GA, *[email protected]
REFERENCES
1) World Health Organization: Global Tuberculosis Report (2018)
2) World Health Organization: Guidelines for treatment of drug-susceptible tuberculosis and patient care (2017)
3) Al-Saeedi M & Al-Hajoj S (2017) Infect. Drug Resist. 10: 333–342
4) Hemsley CM, Luo JX, Andreae CA, Butler CS, Soyer OS & Titball RW (2014) Antimicrob. Agents Chemother. 58: 5775–5783
5) Choudhary E, Sharma R, Kumar Y, Agarwal N (2019). Frontiers in Cellular and Infection Microbiology. 9:70.
6) Worth CL, Preissner R, Blundell TL (2011). Nucleic acids research. 39:W215-22.
7) biotium.com/product/glomelt-thermal-shift-protein-stability-kit/
8) Bai, N., Roder, H., Dickson, A., & Karanicolas, J. (2019). Scientific reports, 9(1), 2650.
9)www.creativebiomart.net/resource/principle-protocol-x-ray-crystallography
Crystal structure of Mtb RecA (PDB: 4PSV)
Mutation-structure analysis using SDM6Example of Atomwise ligand predictions
Differential Scanning Fluorimetry will be
used to screen which compounds bind to
Mtb RecA7
Isothermal titration calorimetry will be
used to measure binding affinity8
X-Ray crystallography will be used to determine
structure of compound-RecA complex9
Surface Plasmon Resonance will be used
to measure binding affinity8
Example plasmid that will be used to produce
Mtb RecA in E. coli
Compounds predicted by Atomwise to inhibit
the ATP binding spot of Mtb RecA (example
compounds)