Inhibiting Pneumococcal Surface Antigen A (PsaA) with Small Molecules Discovered Through Virtual Screening: Steps Toward Validating A Potential Target for Streptococcus pneumoniae.

The pneumococcal surface antigen A (PsaA) metal transporter protein provides manganese to bacterial cells. X-ray crystal structures of PsaA, in both closed (Mn bound) and open (metal free) conformations, were explored with virtual screening to identify potential inhibitors of manganese transport. We pursued three strategies for inhibition: 1) targeting a cavity close to the bound Mn to keep the metal in-place; 2) targeting the metal-free Mn site to prevent metal uptake; and 3) targeting a potentially druggable allosteric site involving loops that translate between the conformations. Tiered assays were used to test the resulting 170 acquired hits: a) assay 1 tested the compounds' growth inhibition of the TIGR4 S. pneumoniae strain (ΔpsaA mutant control), yielding 80 compounds (MIC ≤ 250 μM); b) assay 2 tested if addition of 20 μM Mn to inhibited cell cultures restored growth, yielding 21 compounds; and c) assay 3 confirmed that restored bacterial growth was Mn concentration dependent, as was restoration of ΔpsaA growth, yielding 12 compounds with MICs of 125 μM or greater. It may be possible for a small molecule to inhibit PsaA, but we have not yet identified a compound with exemplary properties.