Figure 2.

Scheme for the development of targeted ester prodrugs. (A) The in vitro 2TA–terbium luminescence assay is used to identify alcohol moieties that modulate esterase activity in desired cell lysates. (B) Candidate carboxylate compounds are unearthed through literature mining, machine learning, or other approaches. (C) Carboxylate compounds are condensed with the alcohol compounds of interest and their activity against target and off-target cell lines assessed using a viability assay. (D) Genomic and bioinformatic analyses are used to determine which esterase(s) present in the target strain(s) are responsible for the observed cleavage. These data further suggest which strains are susceptible to the compounds and which strains are not. Additional viability assays can then be employed to confirm conclusions.