Abstract

A high throughput protein workflow has been developed at LifeArc to advance small molecule drug discovery projects from target and construct selection through fragment screening to hit validation. Antimicrobial drug resistance is a public health crisis facing the world and novel antimicrobial therapies are required to help address this challenge. We will show how we have applied our workflow to early stage antimicrobial drug discovery work, progressing from lists of essential genes to structurally-enabled validated hits ready for medicinal chemistry progression. The workflow typically begins with selection of potential drug targets from functional genomic data (for example bacterial gene essentiality data), then progresses to high throughput cloning, protein expression testing and quality control, followed by fragment screening and final target selection. Protein constructs that pass quality control steps are also used for biochemical assays and structural biology to further validate fragment hits. High throughput approaches throughout allow the efficient parallel testing of different protein constructs, comparison of different drug targets for ligandability and generate large fragment binding data sets to empower decision making & provide FBDD starting points. This approach has enabled target prioritisation from a long list of potential antimicrobial targets and has generated hits to kick start our antimicrobial small molecule drug discovery projects, however this workflow could equally be used in any project in which there are multiple targets of interest, including across target classes or pathways.