6 research outputs found
Optimization of CoaD Inhibitors against Gram-Negative Organisms through Targeted Metabolomics
Drug-resistant Gram-negative bacteria
are of increasing concern worldwide. Novel antibiotics are needed,
but their development is complicated by the requirement to simultaneously
optimize molecules for target affinity and cellular potency, which
can result in divergent structure–activity relationships (SARs).
These challenges were exemplified during our attempts to optimize
inhibitors of the bacterial enzyme CoaD originally identified through
a biochemical screen. To facilitate lead optimization, we developed
mass spectroscopy assays based on the hypothesis that levels of CoA
metabolites would reflect the cellular enzymatic activity of CoaD.
Using these methods, we were able to monitor the effects of cellular
enzyme inhibition at compound concentrations up to 100-fold below
the minimum inhibitory concentration (MIC), a common metric of growth
inhibition. Furthermore, we generated a panel of efflux pump mutants
to dissect the susceptibility of a representative CoaD inhibitor to
efflux. These approaches allowed for a nuanced understanding of the
permeability and efflux liabilities of the series and helped guide
optimization efforts to achieve measurable MICs against wild-type <i>E. coli</i>
Discovery and Optimization of Phosphopantetheine Adenylyltransferase Inhibitors with Gram-Negative Antibacterial Activity
In the preceding
manuscript [Moreau et al. 2018, 10.1021/acs.jmedchem.7b01691]
we described a successful fragment-based lead discovery
(FBLD) strategy for discovery of bacterial phosphopantetheine adenylyltransferase
inhibitors (PPAT, CoaD). Following several rounds of optimization
two promising lead compounds were identified: triazolopyrimidinone <b>3</b> and 4-azabenzimidazole <b>4</b>. Here we disclose
our efforts to further optimize these two leads for on-target potency
and Gram-negative cellular activity. Enabled by a robust X-ray crystallography
system, our structure-based inhibitor design approach delivered compounds
with biochemical potencies 4–5 orders of magnitude greater
than their respective fragment starting points. Additional optimization
was guided by observations on bacterial permeability and physicochemical
properties, which ultimately led to the identification of PPAT inhibitors
with cellular activity against wild-type <i>E. coli.</i
Discovery and Optimization of Phosphopantetheine Adenylyltransferase Inhibitors with Gram-Negative Antibacterial Activity
In the preceding
manuscript [Moreau et al. 2018, 10.1021/acs.jmedchem.7b01691]
we described a successful fragment-based lead discovery
(FBLD) strategy for discovery of bacterial phosphopantetheine adenylyltransferase
inhibitors (PPAT, CoaD). Following several rounds of optimization
two promising lead compounds were identified: triazolopyrimidinone <b>3</b> and 4-azabenzimidazole <b>4</b>. Here we disclose
our efforts to further optimize these two leads for on-target potency
and Gram-negative cellular activity. Enabled by a robust X-ray crystallography
system, our structure-based inhibitor design approach delivered compounds
with biochemical potencies 4–5 orders of magnitude greater
than their respective fragment starting points. Additional optimization
was guided by observations on bacterial permeability and physicochemical
properties, which ultimately led to the identification of PPAT inhibitors
with cellular activity against wild-type <i>E. coli.</i
Discovery and Optimization of Phosphopantetheine Adenylyltransferase Inhibitors with Gram-Negative Antibacterial Activity
In the preceding
manuscript [Moreau et al. 2018, 10.1021/acs.jmedchem.7b01691]
we described a successful fragment-based lead discovery
(FBLD) strategy for discovery of bacterial phosphopantetheine adenylyltransferase
inhibitors (PPAT, CoaD). Following several rounds of optimization
two promising lead compounds were identified: triazolopyrimidinone <b>3</b> and 4-azabenzimidazole <b>4</b>. Here we disclose
our efforts to further optimize these two leads for on-target potency
and Gram-negative cellular activity. Enabled by a robust X-ray crystallography
system, our structure-based inhibitor design approach delivered compounds
with biochemical potencies 4–5 orders of magnitude greater
than their respective fragment starting points. Additional optimization
was guided by observations on bacterial permeability and physicochemical
properties, which ultimately led to the identification of PPAT inhibitors
with cellular activity against wild-type <i>E. coli.</i
Fragment-Based Drug Discovery of Inhibitors of Phosphopantetheine Adenylyltransferase from Gram-Negative Bacteria
The
discovery and development of new antibiotics capable of curing
infections due to multidrug-resistant and pandrug-resistant Gram-negative
bacteria are a major challenge with fundamental importance to our
global healthcare system. Part of our broad program at Novartis to
address this urgent, unmet need includes the search for new agents
that inhibit novel bacterial targets. Here we report the discovery
and hit-to-lead optimization of new inhibitors of phosphopantetheine
adenylyltransferase (PPAT) from Gram-negative bacteria. Utilizing
a fragment-based screening approach, we discovered a number of unique
scaffolds capable of interacting with the pantetheine site of <i>E. coli</i> PPAT and inhibiting enzymatic activity, including
triazolopyrimidinone <b>6</b>. Structure-based optimization
resulted in the identification of two lead compounds as selective,
small molecule inhibitors of bacterial PPAT: triazolopyrimidinone <b>53</b> and azabenzimidazole <b>54</b> efficiently inhibited <i>E. coli</i> and <i>P. aeruginosa</i> PPAT and displayed
modest cellular potency against the efflux-deficient <i>E. coli</i> Δ<i>tolC</i> mutant strain
Fragment-Based Drug Discovery of Inhibitors of Phosphopantetheine Adenylyltransferase from Gram-Negative Bacteria
The
discovery and development of new antibiotics capable of curing
infections due to multidrug-resistant and pandrug-resistant Gram-negative
bacteria are a major challenge with fundamental importance to our
global healthcare system. Part of our broad program at Novartis to
address this urgent, unmet need includes the search for new agents
that inhibit novel bacterial targets. Here we report the discovery
and hit-to-lead optimization of new inhibitors of phosphopantetheine
adenylyltransferase (PPAT) from Gram-negative bacteria. Utilizing
a fragment-based screening approach, we discovered a number of unique
scaffolds capable of interacting with the pantetheine site of <i>E. coli</i> PPAT and inhibiting enzymatic activity, including
triazolopyrimidinone <b>6</b>. Structure-based optimization
resulted in the identification of two lead compounds as selective,
small molecule inhibitors of bacterial PPAT: triazolopyrimidinone <b>53</b> and azabenzimidazole <b>54</b> efficiently inhibited <i>E. coli</i> and <i>P. aeruginosa</i> PPAT and displayed
modest cellular potency against the efflux-deficient <i>E. coli</i> Δ<i>tolC</i> mutant strain