Fig 1.
Clinical candidates for Chagas disease.
Fig 2.
a: Traditional endogenous substrate (Lanosterol) and CYP51 specific metabolite (FF-MAS); b: Fluorogenic probe substrate BOMCC is metabolised to CHC by CYP51.
Fig 3.
a: Linearity of CHC production (expressed as average rate of fluorescence units (AFU)) with increasing T. cruzi CYP51 enzyme concentration (96 well format; average points (n = 3) with error bars); b: Michaelis-Menten Kinetic determination (96 well format; average points (n = 3) with error bars).
Fig 4.
a: The effect of solvent on T. cruzi CYP51 activity; b: Cytochrome C reductase activity of recombinant preparations.
Table 1.
IC50 Determination of a test set of compounds.
Fig 5.
a: T. cruzi CYP51 inhibition plotted against T. cruzi activity for compounds from the different chemical series (coloured green), in addition to 16 known inhibitors of human CYP51 used as a reference set to define the correlation curve (coloured red). T. cruzi active series within the dotted box are prioritised for further medicinal chemistry; b: T. cruzi CYP51 inhibition plotted against T. cruzi activity for series one (green), and series two (blue) compared with the human CYP51 training set (red); c: T. cruzi CYP51 inhibition plotted against T. cruzi activity for a third series (yellow) compared with the human CYP51 training set (red). pEC50 or pIC50 is the −log of EC50 or IC50 respectively in M.
Fig 6.
Chemical moieties associated with CYP inhibition.
Fig 7.
Trends of control 1 (closed circles, solvent only control, 0% inhibition), control 2 (open circles, reaction in absence of bactosomes, 100% inhibition) and Z’ (grey squares) of representative high-throughput screening runs of T. cruzi CYP51 FLINT (a) and cytochrome c reductase absorbance (b) assays in 384 well format.
Error bars are the standard deviations of 16 replicates for each control and plate.