Escalating multidrug resistance (MDR) in Neisseria gonorrhoeae (Ng) and the absence of a vaccine underscore the urgent need for new therapeutics. We established an integrated programme combining phenotypic screening with target-based anti-virulence approaches, enabling parallel generation of bactericidal leads and mechanistically defined inhibitors of host–pathogen interactions.
High-throughput phenotypic screens were conducted across multiple libraries containing natural products and their synthetic and semi-synthetic derivatives. We identified 20 compounds active against susceptible and MDR Ng, with minimum inhibitory and bactericidal concentrations (MIC/MBC) ranging from 0.7–55 µg/mL across five strains. Thirteen compounds showed consistent MICs across four MDR reference strains and similar activity in wild-type and ΔmtrE Ng, indicating they are not subject to MtrE-mediated efflux. In cervical epithelial assays, 10/20 compounds were non-toxic at active doses, and 7/20 reduced colony-forming units (CFU) by 77–96% at ½ MIC in an infection model without detectable cytotoxicity. Based on potency, bactericidal activity, reduced susceptibility to efflux, and host-cell safety, we prioritised six lead compounds for further development.
In parallel, we established a multiplatform, target-based discovery pipeline focused on essential virulence functions, integrating structure-based docking, deep learning–guided prioritisation, and surface plasmon resonance (SPR)-based biophysical validation to identify druggable targets and inhibitors. Using this approach, we targeted N. gonorrhoeae glycan-binding proteins, including NHBA and TbpA, which mediate host interactions involved in adherence, nutrient acquisition, and immune evasion. To identify inhibitors, we performed in silico screening of commercially available libraries followed by SPR validation to prioritise leads. In a pilot screen of a 2,400-compound FDA-approved library enriched for clinically advanced molecules, we identified five candidates that inhibit glycan-dependent interactions, including TbpA-mediated adherence and transferrin binding.
In summary, this integrated platform delivers both bactericidal leads and inhibitors of host–pathogen interactions, providing complementary paths toward new therapeutics for MDR N. gonorrhoeae and other AMR pathogens.