Nanomole-scaled high-throughput chemistry plus direct bioautography on the same chromatography plate for drug discovery

Abstract

The powerful fusion of on-surface synthesis and effect-directed analysis was introduced as novel tool for synthetic drug discovery, all on the same high-performance thin-layer chromatography plate. Precise automated sample application allowed both, high-throughput chemistry of 60 reactions at once and reaction miniaturization down to the 15-nmol scale. The antibiotic activity of all on-surface synthesized compounds was evaluated on the same surface via the Gram-positive Bacillus subtilis bioassay. For one product, synthesis (reaction, purification and identification) took 5.3 min and semi-quantitative biological evaluation took 2.8 min. Out of 60 on-surface reactions 10 products (17%) were identified to be more active than a well-known antibiotic reference. The concept was transferred to the Gram-negative Aliivibrio fischeri bioassay. For the first time, a new analytical platform was shown for a streamlined workflow at the most miniaturized scale from synthesis, purification, identification and quantification to semi-quantitative biological activity evaluation (all on the same chromatography plate).

Keywords: 2-Amino-3-cyano-4H-pyran; Antibiotic; Antimicrobial; Drug development; Miniaturized on-surface synthesis; Planar chromatography.