Reengineering Vancomycin to Combat Bacterial Resistance. Matthew Giletto September 18, 2013 CEM 958

Reengineering Vancomycin to Combat Bacterial Resistance Matthew Giletto September 18, 2013 CEM 958

Overview Why bacterial resistance to antibiotics is an important area of research Review the history of vancomycin, its structural elucidation and mechanism of action Track the development of bacterial resistance to vancomycin Examine SAR work on vancomycin Learn how vancomycin has been assembled in the laboratory and propose how this knowledge may let us build better vancomycin(s)

Bacterial resistance to antibiotics Meticillin resistant Staphylococcus aureus, MRSA, killed 19,000 people (2005) invasively effected 94,000 in the US (2005) 3-4 billion dollars (2005) Vancomycin was last line defense against multidrug resistant pathogens Vancomycin resistant S. aureus, VRSA Vancomycin resistant Enterococci, VRE Resistance is acquired as a result of gene transfer from nonpathogens to pathogens and between pathogens Walsh, C. T.; Fischbach M. A. Sci. Am., 2009, 301, 44. Klevens, R. M. et al. J. Am. Med. Assoc., 2007; 298, 1763. Wengel, L. et. al. Science; 2003; 302, 1569. Neu, H.C. Science, 1992, 257, 1064. Leclercq, R. et. al. N. Eng. J. Med., 1988, 319, 157.

Solution To use organic synthesis (total synthesis, semisynthesis and catalysis) as a tool to solve problems in diverse areas of science (chemistry, biology, medicine) that are not solvable with other methods

Classes of antibiotics active against Gram Positive pathogens

X-ray Structure of CDP-I Bardsley, B., Williams, D. H.; Angew. Chemie. Int. Ed.; 1999; 38; 1172. Williams, D. H.; Williamson, M. P.; J. Am. Chem. Soc.; 1981; 103; 6580. Harris, C. M.; Harris, T. M.; J. Am. Chem. Soc.; 1982; 104; 4293. Williams, D. H. et. al.; Nature; 1978; 271; 223. Marshall, F. J.; J. Med. Chem.; 1965; 8; 18.

Rearrangement to CDP-I Boger, D. L. et. al. J. Am. Chem. Soc., 1998, 120, 8920. Harris, C. M.; Harris, T. M. J. Am. Chem. Soc., 1982, 104, 4293.

Key noe s of CDP-I and vancomycin Nitanai, Y. et. al. J. Mol. Biol., 2009, 385, 1422. Loll, P. J. J. Am. Chem. Soc., 1997, 119, 1516. Williams, D. H.; Williamson, M. P.. J. Am. Chem. Soc., 1981, 103, 6580.

Vancomycin inhibits cell wall synthesis at transglycosylation Schaefer: D-[1-13 C]-ala incorporation exclusive to cell wall precursors AND quantitatively detectable in solid state NMR % D- [1-13 C]-ala-D-[1-13 C]-ala in growing Enterococci is 24 % D- [1-13 C]-ala-D-[1-13 C]-ala 45 min after 25 mg/ml vancomycin doubles to 48 Diagnostic of accumulation of cell wall precursors in cytoplasm Schaefer, J. et. al. Biochemistry, 2013, 52, 3405. Schaefer, J. et. al. J. Mol. Biol., 2009, 392, 1178. Schaefer, J. et. al. J. Mol. Biol., 2006, 357, 1253. Kricheldorf, H. R.; Muller, D. Macromolecules, 1983, 16, 615.

Bacterial cell wall synthesis: Phase 1 Schaefer, J. et. al. Biochemistry, 2013, 52, 3405. Schaefer, J. et. al. J. Mol. Biol., 2009, 392, 1253. Kahne, D. et. al. Chem. Rev., 2005, 105, 425.

Installation of D-asp bridge in Enterococci Bellias, S. et. al. J. Biol. Chem., 2006, 281, 11586.

Kahne, D. et. al. Chem. Rev., 2005, 105, 425. Phase 2

Kahne, D. et. al. Chem. Rev., 2005, 105, 425.

Phase III Step 1:transglycosylation Kahne, D. et. al. Chem. Rev., 2005, 105, 425.

Binding model in susceptible bacteria Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 1284. Williams, D. H.; Bardsley, B. Angew. Chemie. Int. Ed., 1999, 38, 1172. Williams, D. H. et. al. J. Am. Chem. Soc., 1983, 105, 1332. Perkins, H.R.; Nieto, M. Biochem. J., 1971, 123, 789.

Binding model in resistant bacteria Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 1284. Williams, D. H.; Bardsley, B. Angew. Chemie. Int. Ed., 1999, 38, 1172. Williams, D. H. et. al. J. Am. Chem. Soc., 1983, 105, 1332. Perkins, H.R.; Nieto, M. Biochem. J., 1971, 123, 789.

Potential dual binding capacity of amidines Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 1284. Boger, D. L.; Crowley, B. M. J. Am. Chem. Soc., 2006, 128, 2885.

Activating the mechanism of resistance Wright G. D. et. al. Nature Chemical Biology, 2010, 6, 327.

Probing the SAR for possible solutions to bacterial resistance

Des-leucyl vancomycin series Kahne, D. et. al. Chem. Rev., 2005, 105, 425. Kahne, D. et. al.; Science, 1999; 284, 507. Williams, D H. et. al J. Antibiot., 1995, 48, 805.

Des-leucyl vancomycin series Kahne, D. et. al. Chem. Rev., 2005, 105, 425. Kahne, D. et. al.; Science, 1999; 284, 507. Williams, D H. et. al J. Antibiot., 1995, 48, 805.

Des-leucyl chlorobiphenyl vancomycin series Schaefer, J. et. al. J. Mol. Biol., 2009, 392, 1253. Kahne, D. et. al. Chem. Rev., 2005, 105, 425. Kahne, D. et. al. Science, 1999, 284, 507.

Des-leucyl chlorobiphenyl vancomycin series The complexity of the peptide portion of vancomycin makes it virtually impossible to reengineer the peptide backbone to include new contacts to the modified substrate. D. Kahne Schaefer, J. et. al. J. Mol. Biol., 2009, 392, 1253. Kahne, D. et. al. Chem. Rev., 2005, 105, 425. Kahne, D. et. al. Science, 1999, 284, 507.

Oritavancin Zhanel, G. G. et. al. Drugs, 2010, 70, 859. Schaefer, J. Biochemistry, 2008, 47, 10155. Allen, N. et. al. J. Antibiot., 1997, 50, 677.

Oritavancin inhibits transpeptidation Schaefer, J. et. al. J. Mol. Biol., 2009, 392, 1253.

Site-Selective bromination of vancomycin Miller, S.J.; Pathak, T.P. J. Am. Chem. Soc., 2012, 134, 6120.

Proposed Binding Model Miller, S.J.; Pathak, T.P. J. Am. Chem. Soc., 2012, 134, 6120.

Modifications external to binding site Modifying carbohydrate = new mechanism Catalysis Schaefer, J. et. al. Biochemistry, 2013, 52, 3405. Miller, S.J.; Pathak, T.P. J. Am. Chem. Soc., 2012, 134, 6120. Schaefer, J. et. al. J. Mol. Biol., 2009, 392, 1253.

Redesigning vancomycin Total syntheses: Nicolaou, Evans, Boger Nicolaou, K. C. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2708. Evans, D. A. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2700. Boger, D. L. et. al. J. Am. Chem. Soc., 1999, 121, 3226. Smith, G. G. et. al. J. Org. Chem., 1983, 48, 5368.

Retrosynthetic analysis Kahne, D. et. al. J. Am. Chem. Soc., 1998, 120, 11014. Nicolaou, K. C. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2708. Evans, D. A. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2700. Boger, D. L. et. al. J. Am. Chem. Soc., 1999, 121, 3226.

Retrosynthesis of Eastern Hemisphere Nicolaou, K. C. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2708. Evans, D. A. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2700. Boger, D. L. et. al. J. Am. Chem. Soc., 1999, 121, 3226.

Retrosynthesis of Western Hemisphere Nicolaou, K. C. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2708. Evans, D. A. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2700. Boger, D. L. et. al. J. Am. Chem. Soc., 1999, 121, 3226.

The Nicolaou retrosynthetic approach Nicolaou, K. C. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2708.

Synthesizing the AB ring atropisomer Nicolaou, K. C. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2708.

Synthesizing the CD macrocycle Nicolaou, K. C. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2708.

Synthesizing the DE macrocycle Nicolaou, K. C. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2708.

Learning from the Nicolaou approach Nicolaou, K. C. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2708.

Evans Retro of the Western Hemisphere Evans, D. A. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2700.

Synthesizing of AB macrocycle Evans, D. A. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2700.

Synthesizing the CD macrocycle Evans, D. A. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2700.

Preparing for the AB ring equilibration Evans, D. A. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2700.

Equilibrating the AB macrocycle Evans, D. A. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2700. Evans, D. A. J. Am. Chem. Soc., 1993, 115, 6426.

Synthesizing the DE macrocycle Evans, D. A. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2700.

Evans synthesis Evans, D. A. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2700.

Nicolaou versus Evans Nicolaou, K. C. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2708. Evans, D. A. et. al. Angew. Chemie. Int. Ed., 1998, 37, 2700.

The Boger strategy: Equilibration Boger, D. L. et. al. J. Am. Chem. Soc., 1998, 120, 8920.

The Boger synthesis of vancomycin amidine aglycon Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 1284.

Incorporating the A ring Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 1284.

Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 1284.

Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 1284.

Completion Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 1284.

Vancomycin aglycon vs vancomycin amidine aglycon Boger, D. L.. et. al. J. Am. Chem. Soc., 2012, 134, 1284.

Dual binding capacity of amidines Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 1284.

Analogs and factors influencing binding Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 1284. Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 8790.

An optimized analog: Amidine Oritavancin

Proposed traditional route Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 1284.

Amidine Oritavancin Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 1284. Nicolaou, K. C. et. al. Chem. Eur. J., 1999, 5, 2648.

Amidine Oritavancin Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 1284. Nicolaou, K. C. et. al. Chem. Eur. J., 1999, 5, 2648.

Boger, D. L. et. al. J. Am. Chem. Soc.; 2012; 134, 1284. Nicolaou, K. C. et. al. Chem. Eur. J., 1999, 5, 2648.

Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 1284. Nicolaou, K. C. et. al. Chem. Eur. J., 1999, 5, 2648.

Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 1284. Nicolaou, K. C. et. al. Chem. Eur. J., 1999, 5, 2648.

Amidine Oritavancin Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 1284. Nicolaou, K. C. et. al. Chem. Eur. J., 1999, 5, 2648.

Amidine Oritavancin

Proposed peptide catalytic route to Amidine Oritavancin Boger, D. L. et. al. J. Am. Chem. Soc., 2012, 134, 1284.

Miller and Ley Miller, S.J.; Pathak, T.P. J. Am. Chem. Soc., 2012, 134, 6120. Ley, S. V. et. al. J. Chem. Soc. Perkin Trans. I, 2001, 358.

A combined approach Miller, S.J.; Pathak, T.P. J. Am. Chem. Soc., 2012, 134, 6120. Ley, S. V. et. al. J. Chem. Soc. Perkin Trans. I, 2001, 358.

A peptidomimetic Lawessons reagent Miller, S.J.; Pathak, T.P. J. Am. Chem. Soc., 2012, 134, 6120. Joullie, M. et. al. J. Am. Chem. Soc.; 2002; 124; 520. Ley, S. V. et. al. J. Chem. Soc. Perkin Trans. I, 2001, 358.

Putative binding model Miller, S.J.; Pathak, T.P. J. Am. Chem. Soc., 2012, 134, 6120. Ley, S. V. et. al. J. Chem. Soc. Perkin Trans. I, 2001, 358.

Conclusions Important scientific problems can be better understood and solved with the tools of organic synthesis Specifically the problem of inevitable evolution of bacterial resistance to antibiotics can be countered in ways that only organic synthesis could accomplish, restoring our ability to combat deadly and otherwise untreatable diseases

Thanks Dr. Tepe and current group members Nicole Hewlett Travis Bethel Greg Patten Jacob Ludwig Dr. Huang The audience Support of Holeigh, friends, and family