Bacterial defiance as a form of prodrug failure

Brandon Alexander Holt; Gabriel A. Kwong

All control samples containing either TEV-specific prodrug alone or Aprotinin inhibitor did not 6 significantly reduce bacteria load (Fig. 1E, F, Table S1 ). Our results showed that AMP drug-lock complexes are inert and lack cytotoxic activity until activation by protease activity. The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/556951 doi: bioRxiv preprint or OmpT expressed on the surface of E. coli (right) against fluorescently labeled hairpin prodrugs (blue lines) or hairpin prodrugs only control (grey lines). Shading represents standard deviation (n = 3). All cleavage assays plotted as fold change in RFU from initial time point. (E) Bacteria viability assay quantifying drug toxicity relative to untreated bacteria control (blue bar). Positive control for AMP toxicity (black bar). Negative control for locked AMP or TEV protease alone (tan 5 bars). Positive control for TEV protease with locked AMP (substrate: ENLYFQ|G, specific to TEV protease) (red bar). Negative control for locked AMP (substrate: RRSRRV, specific to OmpT) with Representative images of bacterial plates used to quantify viability with schematic legend (scale bar = 4 mm). * < 0.05, ** < 0.01, *** < 0.001, and **** < 0.0001.