Discovery of Inhibitors of Shiga Toxin Type 2 by On-Plate Generation and Screening of a Focused Compound Library

• Published in: Angewandte Chemie International Edition 2014-02, Vol.53 (6), p.1510-1515
• Main Authors: Dasgupta, Somnath, Kitov, Pavel I., Sadowska, Joanna M., Bundle, David R.
• Format: Article
• Language: English
• Subjects: Affinity; Arrays; Conjugates; drug discovery; Enzyme Inhibitors - chemistry; Enzyme Inhibitors - metabolism; Enzyme-Linked Immunosorbent Assay; Escherichia coli - metabolism; glycoarrays; haemolytic uremic syndrome; Inhibitors; Libraries; Ligands; Protein Binding; Screening; Shiga Toxin 2 - antagonists & inhibitors; Shiga Toxin 2 - metabolism; Shiga toxins; Small Molecule Libraries - chemistry; Small Molecule Libraries - metabolism; Toxins
• ISSN: 1433-7851; 1521-3773
• DOI: 10.1002/anie.201309436

A new microtiter‐plate‐based method for the rapid generation and evaluation of focused compound libraries was developed and applied to screening ligand analogues for the E. coli Shiga‐like toxin Stx2a. The method is general, it mitigates the masking of intrinsic affinity gains by multivalency and enables the discovery of potential hits when starting from ligands that exhibit extremely low affinity with proteins that depend on multivalency for their function. Target the toxin: An array of modified ligands was generated after coating microtiter plates with a common precursor containing a reactive functionality. Physically adsorbed ligand–polymer conjugates tolerate a variety of reaction conditions and support instant evaluation of the library by ELISA, as well as subsequent iterative cycles of modification and evaluation. High‐affinity ligands for E. coli Shiga toxin type 2 were discovered by using this method.